Package: metagene
Modified: 18 september, 2015
Compiled: Wed Oct 10 20:10:51 2018
License: Artistic-2.0 | file LICENSE
This package produces metagene-like plots to compare the behavior of DNA-interacting proteins at selected groups of features. A typical analysis can be done in viscinity of transcription start sites (TSS) of genes or at any regions of interest (such as enhancers). Multiple combinations of group of features and/or group of bam files can be compared in a single analysis. Bootstraping analysis is used to compare the groups and locate regions with statistically different enrichment profiles. In order to increase the sensitivity of the analysis, alignment data is used instead of peaks produced with peak callers (i.e.: MACS2 or PICS). The metagene package uses bootstrap to obtain a better estimation of the mean enrichment and the confidence interval for every group of samples.
This vignette will introduce all the main features of the metagene package.
library(metagene)
There is no hard limit in the number of BAM files that can be included in an
analysis (but with too many BAM files, memory may become an issue). BAM files
must be indexed. For instance, if you use a file names file.bam
, a file
named file.bam.bai
or file.bai
must be present in the same directory.
The path (relative or absolute) to the BAM files must be in a vector:
bam_files <- get_demo_bam_files()
bam_files
## [1] "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep1.bam"
## [2] "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep2.bam"
## [3] "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep1.bam"
## [4] "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep2.bam"
## [5] "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/ctrl.bam"
For this demo, we have 2 samples (each with 2 replicates). It is also possible to use a named vector to add your own names to each BAM files:
named_bam_files <- bam_files
names(named_bam_files) <- letters[seq_along(bam_files)]
named_bam_files
## a
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep1.bam"
## b
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep2.bam"
## c
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep1.bam"
## d
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep2.bam"
## e
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/ctrl.bam"
Using named BAM files can simplify the use of the metagene helper functions and the creation of the design.
To compare custom regions of interest, it is possible to use a list of one or more BED files.
regions <- get_demo_regions()
regions
## [1] "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/list1.bed"
## [2] "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/list2.bed"
The name of the files (without the extension) will be used to name each groups.
metagene
also support the
narrowPeak
and the broadPeak.
As an alternative to a list of BED files, GRanges
or GRangesList
objects
can be used.
Some common datasets are already available with the metagene
package:
promoters_hg19
promoters_hg18
promoters_mm10
promoters_mm9
data(promoters_hg19)
promoters_hg19
## GRanges object with 23056 ranges and 1 metadata column:
## seqnames ranges strand | gene_id
## <Rle> <IRanges> <Rle> | <character>
## 1 chr19 58873215-58875214 - | 1
## 10 chr8 18247755-18249754 + | 10
## 100 chr20 43279377-43281376 - | 100
## 1000 chr18 25756446-25758445 - | 1000
## 10000 chr1 244005887-244007886 - | 10000
## ... ... ... ... . ...
## 9991 chr9 115094945-115096944 - | 9991
## 9992 chr21 35735323-35737322 + | 9992
## 9993 chr22 19108968-19110967 - | 9993
## 9994 chr6 90538619-90540618 + | 9994
## 9997 chr22 50963906-50965905 - | 9997
## -------
## seqinfo: 93 sequences (1 circular) from hg19 genome
For more details about each datasets, please refer to their documentation
(i.e.:?promoters_hg19
).
A design group contains a set of BAM files that, when put together, represent a logical analysis. Furthermore, a design group contains the relationship between every BAM files present. Samples (with or without replicates) and controls can be assigned to a same design group. There can be as many groups as necessary. A BAM file can be assigned to more than one group.
To represent the relationship between every BAM files, design groups must have the following columns:
There is two possible way to create design groups, by reading a file or by directly creating a design object in R.
Design groups can be loaded into the metagene package by using a text file. As the relationship between BAM files as to be specified, the following columns are mandatory:
The file must also contain a header. It is recommanded to use Samples
for the
name of the first column, but the value is not checked. The other columns in
the design file will be used for naming design groups, and must be unique.
fileDesign <- system.file("extdata/design.txt", package="metagene")
design <- read.table(fileDesign, header=TRUE, stringsAsFactors=FALSE)
design$Samples <- paste(system.file("extdata", package="metagene"),
design$Samples, sep="/")
kable(design)
Samples | align1 | align2 |
---|---|---|
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep1.bam | 1 | 0 |
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep2.bam | 1 | 0 |
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep1.bam | 0 | 1 |
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep2.bam | 0 | 1 |
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/ctrl.bam | 2 | 2 |
It is not obligatory to use a design file, you can create the design
data.frame
using your prefered method (as long as the restrictions on the
values mentioned previously are respected).
For instance, the previous design data.frame could have been create directly in R:
design <- data.frame(Samples = c("align1_rep1.bam", "align1_rep2.bam",
"align2_rep1.bam", "align2_rep2.bam", "ctrl.bam"),
align1 = c(1,1,0,0,2), align2 = c(0,0,1,1,2))
design$Samples <- paste0(system.file("extdata", package="metagene"), "/",
design$Samples)
kable(design)
Samples | align1 | align2 |
---|---|---|
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep1.bam | 1 | 0 |
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep2.bam | 1 | 0 |
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep1.bam | 0 | 1 |
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep2.bam | 0 | 1 |
/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/ctrl.bam | 2 | 2 |
A typical metagene analysis will consist steps:
A minimal metagene analysis can be performed in 2 steps:
new
function).plot
regions <- get_demo_regions()
bam_files <- get_demo_bam_files()
# Initialization
mg <- metagene$new(regions = regions, bam_files = bam_files)
# Plotting
mg$plot(title = "Demo metagene plot")
## produce data table : ChIP-Seq
## produce data frame : ChIP-Seq
## Plot : ChIP-Seq
As you can see, it is not mandatory to explicitly call each step of the
metagene analysis. For instance, in the previous example, the plot
function
call the other steps automatically with default values (the next section will
describe the steps in more details).
In this specific case, the plot is messy since by default metagene will produce a curve for each possible combinations of BAM file and regions. Since we have 5 BAM files and 2 regions, this gives us 10 curves.
If we want more control on how every step of the analysis are performed, we have to call each functions directly.
In order to fully control every step of a metagene analysis, it is important to understand how a complete analysis is performed. If we are satisfied with the default values, it is not mandatory to explicitly call every step (as was shown in the previous section).
During this step, the coverages for every regions specified are extracted from
every BAM files. More specifically, a new GRanges
is created by combining
all the regions specified with the regions
param of the new
function.
regions <- get_demo_regions()
bam_files <- get_demo_bam_files()
mg <- metagene$new(regions = regions, bam_files = bam_files)
To produce the table, coverages (produced from Genomics regions (.BED),
Alignment Files (.BAM) and Design Sheet) was treated for noise removal
and normalized. Furthermore, to reduce the computation time during the
following steps, the positions are also binned. Regions, designs, bins,
associated values and orientation of strands are pulled into a data.table
called ‘table’ and accessible thanks to the getter get_table
.
We can control the size of the bins with the bin_count
argument. By
default, a bin_count
of 100 will be used during this step.
mg$produce_table()
## produce data table : ChIP-Seq
We can also use the design we produced earlier to remove background signal and combine replicates:
mg$produce_table(design = design)
## produce data table : ChIP-Seq
data.frame
The metagene plot are produced using the ggplot2
package, which require a
data.frame
as input. During this step, the values of the ribbon are
calculated. Metagene uses “bootstrap” to obtain a better estimation of the
mean of enrichment for every positions in each groups.
mg$produce_data_frame()
## produce data frame : ChIP-Seq
During this step, metagene will use the data.frame
to plot the calculated
values using ggplot2
. We show a subset of the regions by using the
region_names
and design_names
parameter. The region_names
correspond to
the names of the regions used during the initialization. The design_name
will vary depending if a design was added. If no design was added, this param
correspond to the BAM name or BAM filenames. Otherwise, we have to use the
names of the columns from the design.
mg$plot(region_names = "list1", title = "Demo plot subset")
## Plot : ChIP-Seq
metagene
objectsMultiple getters functions are available to access the data that is stored in a
metagene
object.
get_table
To get the data.table containing regions, designs, bins, values at bins and orientation of strands.
mg <- get_demo_metagene()
mg$produce_table()
## produce data table : ChIP-Seq
mg$get_table()
## region design bin value strand
## 1: list1 align1_rep1 1 0.00 *
## 2: list1 align1_rep1 2 0.75 *
## 3: list1 align1_rep1 3 1.55 *
## 4: list1 align1_rep1 4 1.15 *
## 5: list1 align1_rep1 5 1.25 *
## ---
## 49996: list2 ctrl 96 0.00 *
## 49997: list2 ctrl 97 0.00 *
## 49998: list2 ctrl 98 0.00 *
## 49999: list2 ctrl 99 0.00 *
## 50000: list2 ctrl 100 0.00 *
get_matrices
To get the data.table as matrices (the former data structure)
mg <- get_demo_metagene()
mg$produce_table()
## produce data table : ChIP-Seq
m <- mg$get_matrices()
# m$list1$ctrl$input to access to region 'list1' and 'ctrl' design
get_data_frame
get_data_frame = function(region_names = NULL, design_names = NULL) To get the data.frame containing regions and design
mg <- get_demo_metagene()
mg$produce_table()
## produce data table : ChIP-Seq
mg$produce_data_frame()
## produce data frame : ChIP-Seq
mg$get_data_frame()
## region design bin value strand qinf qsup
## 1 list1 align1_rep1 1 0.449 * 0.39664250 0.51437125
## 2 list1 align1_rep1 2 0.552 * 0.50846500 0.61749375
## 3 list1 align1_rep1 3 0.609 * 0.57691875 0.65218000
## 4 list1 align1_rep1 4 0.534 * 0.49703875 0.58920250
## 5 list1 align1_rep1 5 0.522 * 0.47645375 0.55863750
## 6 list1 align1_rep1 6 0.589 * 0.54526500 0.63079500
## 7 list1 align1_rep1 7 0.704 * 0.61347625 0.79250750
## 8 list1 align1_rep1 8 0.591 * 0.55181625 0.63729625
## 9 list1 align1_rep1 9 0.562 * 0.50106625 0.62043000
## 10 list1 align1_rep1 10 0.488 * 0.43104875 0.54088250
## 11 list1 align1_rep1 11 0.614 * 0.54335750 0.68286625
## 12 list1 align1_rep1 12 0.745 * 0.67426125 0.88109000
## 13 list1 align1_rep1 13 0.772 * 0.68679375 0.86025375
## 14 list1 align1_rep1 14 0.659 * 0.57884750 0.73050250
## 15 list1 align1_rep1 15 0.481 * 0.42830125 0.54616875
## 16 list1 align1_rep1 16 0.558 * 0.50471250 0.61320250
## 17 list1 align1_rep1 17 0.614 * 0.56193250 0.65519375
## 18 list1 align1_rep1 18 0.708 * 0.65082375 0.75633750
## 19 list1 align1_rep1 19 0.847 * 0.80188250 0.91411375
## 20 list1 align1_rep1 20 0.745 * 0.69940000 0.78628125
## 21 list1 align1_rep1 21 0.731 * 0.68082000 0.77937625
## 22 list1 align1_rep1 22 0.795 * 0.71695125 0.87031000
## 23 list1 align1_rep1 23 0.772 * 0.70670250 0.82391875
## 24 list1 align1_rep1 24 0.866 * 0.79794625 0.92374375
## 25 list1 align1_rep1 25 0.872 * 0.82233875 0.92497250
## 26 list1 align1_rep1 26 0.841 * 0.78724000 0.91310125
## 27 list1 align1_rep1 27 0.808 * 0.71799625 0.88309875
## 28 list1 align1_rep1 28 0.835 * 0.72978375 0.93240250
## 29 list1 align1_rep1 29 0.915 * 0.83936500 0.98256500
## 30 list1 align1_rep1 30 1.123 * 1.02365375 1.25741625
## 31 list1 align1_rep1 31 1.161 * 1.01686375 1.29170500
## 32 list1 align1_rep1 32 1.122 * 0.96641250 1.25926625
## 33 list1 align1_rep1 33 1.301 * 1.20241250 1.46085250
## 34 list1 align1_rep1 34 1.235 * 1.12486750 1.32650000
## 35 list1 align1_rep1 35 1.285 * 1.20336250 1.35660625
## 36 list1 align1_rep1 36 1.497 * 1.43662875 1.58848250
## 37 list1 align1_rep1 37 1.480 * 1.37057500 1.59496750
## 38 list1 align1_rep1 38 1.433 * 1.25767375 1.55408250
## 39 list1 align1_rep1 39 1.234 * 1.10148750 1.33955625
## 40 list1 align1_rep1 40 1.227 * 1.16291875 1.30427000
## 41 list1 align1_rep1 41 1.302 * 1.20409125 1.38735750
## 42 list1 align1_rep1 42 1.267 * 1.21169250 1.33171500
## 43 list1 align1_rep1 43 1.584 * 1.52268500 1.69705750
## 44 list1 align1_rep1 44 1.602 * 1.52082625 1.67415375
## 45 list1 align1_rep1 45 1.727 * 1.62622500 1.81776375
## 46 list1 align1_rep1 46 2.020 * 1.93262000 2.10656375
## 47 list1 align1_rep1 47 2.120 * 2.01315500 2.22835875
## 48 list1 align1_rep1 48 2.044 * 1.92662000 2.14018375
## 49 list1 align1_rep1 49 2.196 * 2.11249375 2.27402000
## 50 list1 align1_rep1 50 2.340 * 2.24409750 2.44725125
## 51 list1 align1_rep1 51 2.367 * 2.27289375 2.49698750
## 52 list1 align1_rep1 52 2.374 * 2.28761500 2.51700625
## 53 list1 align1_rep1 53 2.086 * 1.96582125 2.19082000
## 54 list1 align1_rep1 54 2.053 * 1.95615375 2.12298500
## 55 list1 align1_rep1 55 2.126 * 1.99515000 2.24243125
## 56 list1 align1_rep1 56 1.710 * 1.62754375 1.80812750
## 57 list1 align1_rep1 57 1.429 * 1.33481000 1.53173875
## 58 list1 align1_rep1 58 1.425 * 1.34745625 1.48506250
## 59 list1 align1_rep1 59 1.618 * 1.50523375 1.69377000
## 60 list1 align1_rep1 60 1.707 * 1.55877625 1.83512125
## 61 list1 align1_rep1 61 1.716 * 1.63249000 1.84332000
## 62 list1 align1_rep1 62 1.699 * 1.58482125 1.79187250
## 63 list1 align1_rep1 63 1.383 * 1.27449375 1.50234750
## 64 list1 align1_rep1 64 1.316 * 1.23834250 1.40915500
## 65 list1 align1_rep1 65 1.293 * 1.20344625 1.36353750
## 66 list1 align1_rep1 66 1.136 * 1.06626625 1.20390500
## 67 list1 align1_rep1 67 1.095 * 1.01749375 1.16033750
## 68 list1 align1_rep1 68 1.028 * 0.96446000 1.09408875
## 69 list1 align1_rep1 69 0.895 * 0.82624250 0.94574500
## 70 list1 align1_rep1 70 0.825 * 0.77847875 0.86839500
## 71 list1 align1_rep1 71 0.898 * 0.83558000 0.94753875
## 72 list1 align1_rep1 72 0.856 * 0.80114125 0.91412750
## 73 list1 align1_rep1 73 0.879 * 0.81818000 0.95552625
## 74 list1 align1_rep1 74 0.899 * 0.84374250 0.94736500
## 75 list1 align1_rep1 75 0.958 * 0.90543500 1.04089750
## 76 list1 align1_rep1 76 1.048 * 0.99860750 1.13168250
## 77 list1 align1_rep1 77 0.963 * 0.89608000 1.03102625
## 78 list1 align1_rep1 78 0.941 * 0.89021750 1.02737125
## 79 list1 align1_rep1 79 0.823 * 0.77330750 0.89215875
## 80 list1 align1_rep1 80 0.564 * 0.52933750 0.61821875
## 81 list1 align1_rep1 81 0.486 * 0.43681000 0.54943500
## 82 list1 align1_rep1 82 0.730 * 0.67149375 0.77786375
## 83 list1 align1_rep1 83 0.820 * 0.77333125 0.88140500
## 84 list1 align1_rep1 84 0.711 * 0.66008250 0.75564625
## 85 list1 align1_rep1 85 0.702 * 0.65335000 0.75356000
## 86 list1 align1_rep1 86 0.664 * 0.60122375 0.72229875
## 87 list1 align1_rep1 87 0.508 * 0.47861000 0.55557625
## 88 list1 align1_rep1 88 0.538 * 0.50318625 0.56956000
## 89 list1 align1_rep1 89 0.582 * 0.55019250 0.61583875
## 90 list1 align1_rep1 90 0.701 * 0.64567000 0.74278125
## 91 list1 align1_rep1 91 0.628 * 0.58942000 0.67070750
## 92 list1 align1_rep1 92 0.649 * 0.59497375 0.68878375
## 93 list1 align1_rep1 93 0.900 * 0.84146000 0.97411250
## 94 list1 align1_rep1 94 0.830 * 0.73559000 0.87789375
## 95 list1 align1_rep1 95 0.590 * 0.53793375 0.63935875
## 96 list1 align1_rep1 96 0.557 * 0.52035375 0.62646750
## 97 list1 align1_rep1 97 0.575 * 0.53008500 0.61102250
## 98 list1 align1_rep1 98 0.525 * 0.47606875 0.57591125
## 99 list1 align1_rep1 99 0.404 * 0.36576750 0.44779125
## 100 list1 align1_rep1 100 0.537 * 0.50661750 0.58294250
## 101 list1 align1_rep2 1 0.092 * 0.07350250 0.12106500
## 102 list1 align1_rep2 2 0.161 * 0.09591000 0.21838250
## 103 list1 align1_rep2 3 0.201 * 0.16917875 0.23024750
## 104 list1 align1_rep2 4 0.147 * 0.12611625 0.16845875
## 105 list1 align1_rep2 5 0.077 * 0.06528500 0.08989250
## 106 list1 align1_rep2 6 0.180 * 0.16085000 0.20034750
## 107 list1 align1_rep2 7 0.267 * 0.22891875 0.30386125
## 108 list1 align1_rep2 8 0.233 * 0.19142375 0.27224500
## 109 list1 align1_rep2 9 0.172 * 0.14480250 0.21299875
## 110 list1 align1_rep2 10 0.253 * 0.22450625 0.28107500
## 111 list1 align1_rep2 11 0.258 * 0.22381375 0.28152750
## 112 list1 align1_rep2 12 0.176 * 0.14599500 0.19334375
## 113 list1 align1_rep2 13 0.193 * 0.17232375 0.21844000
## 114 list1 align1_rep2 14 0.216 * 0.18233000 0.25963125
## 115 list1 align1_rep2 15 0.164 * 0.13897625 0.17813250
## 116 list1 align1_rep2 16 0.123 * 0.10263000 0.14723750
## 117 list1 align1_rep2 17 0.110 * 0.09475250 0.12607875
## 118 list1 align1_rep2 18 0.133 * 0.11329000 0.15542750
## 119 list1 align1_rep2 19 0.169 * 0.14758000 0.19208250
## 120 list1 align1_rep2 20 0.265 * 0.22655875 0.30108375
## 121 list1 align1_rep2 21 0.306 * 0.26424250 0.34721250
## 122 list1 align1_rep2 22 0.399 * 0.35207500 0.43852125
## 123 list1 align1_rep2 23 0.326 * 0.28748250 0.35125375
## 124 list1 align1_rep2 24 0.179 * 0.15616500 0.19260375
## 125 list1 align1_rep2 25 0.251 * 0.21733625 0.28334375
## 126 list1 align1_rep2 26 0.369 * 0.32112375 0.40738875
## 127 list1 align1_rep2 27 0.372 * 0.34305000 0.42328125
## 128 list1 align1_rep2 28 0.320 * 0.27550625 0.35814875
## 129 list1 align1_rep2 29 0.254 * 0.23229000 0.28663875
## 130 list1 align1_rep2 30 0.270 * 0.23834500 0.29454000
## 131 list1 align1_rep2 31 0.382 * 0.34080250 0.44905875
## 132 list1 align1_rep2 32 0.402 * 0.34590125 0.45877500
## 133 list1 align1_rep2 33 0.354 * 0.32057875 0.39513875
## 134 list1 align1_rep2 34 0.499 * 0.45867625 0.53634625
## 135 list1 align1_rep2 35 0.635 * 0.57034625 0.68544125
## 136 list1 align1_rep2 36 0.915 * 0.85266000 0.98770625
## 137 list1 align1_rep2 37 1.010 * 0.90439500 1.08239500
## 138 list1 align1_rep2 38 0.811 * 0.75374750 0.90109375
## 139 list1 align1_rep2 39 0.678 * 0.62065375 0.74215250
## 140 list1 align1_rep2 40 0.727 * 0.64854500 0.79240250
## 141 list1 align1_rep2 41 0.769 * 0.70089750 0.85500000
## 142 list1 align1_rep2 42 0.760 * 0.70060750 0.83594250
## 143 list1 align1_rep2 43 0.647 * 0.60155625 0.71114000
## 144 list1 align1_rep2 44 0.849 * 0.79805000 0.89325000
## 145 list1 align1_rep2 45 1.043 * 0.95264625 1.11240875
## 146 list1 align1_rep2 46 1.037 * 0.97010750 1.10665375
## 147 list1 align1_rep2 47 1.105 * 1.04285000 1.15744000
## 148 list1 align1_rep2 48 1.042 * 0.96393750 1.12356625
## 149 list1 align1_rep2 49 0.911 * 0.86087000 0.95743375
## 150 list1 align1_rep2 50 0.962 * 0.88767375 1.04185750
## 151 list1 align1_rep2 51 1.255 * 1.18668000 1.35233750
## 152 list1 align1_rep2 52 1.254 * 1.19826750 1.31491125
## 153 list1 align1_rep2 53 1.109 * 1.04703625 1.17263375
## 154 list1 align1_rep2 54 1.160 * 1.12062000 1.22479875
## 155 list1 align1_rep2 55 1.112 * 1.04065500 1.16981750
## 156 list1 align1_rep2 56 0.912 * 0.82906250 0.98663500
## 157 list1 align1_rep2 57 0.804 * 0.75031375 0.84248625
## 158 list1 align1_rep2 58 0.721 * 0.67970500 0.76491000
## 159 list1 align1_rep2 59 0.542 * 0.50756125 0.57648250
## 160 list1 align1_rep2 60 0.425 * 0.37493250 0.49851750
## 161 list1 align1_rep2 61 0.479 * 0.41120250 0.52491625
## 162 list1 align1_rep2 62 0.729 * 0.68783375 0.78218375
## 163 list1 align1_rep2 63 0.796 * 0.73923875 0.84759500
## 164 list1 align1_rep2 64 0.647 * 0.60979625 0.71828500
## 165 list1 align1_rep2 65 0.603 * 0.55726875 0.65274375
## 166 list1 align1_rep2 66 0.399 * 0.37319875 0.44559125
## 167 list1 align1_rep2 67 0.430 * 0.38225750 0.46142000
## 168 list1 align1_rep2 68 0.498 * 0.46619625 0.53846875
## 169 list1 align1_rep2 69 0.338 * 0.30267000 0.36690500
## 170 list1 align1_rep2 70 0.440 * 0.39858750 0.48770625
## 171 list1 align1_rep2 71 0.454 * 0.42145250 0.49670750
## 172 list1 align1_rep2 72 0.445 * 0.41437500 0.47119250
## 173 list1 align1_rep2 73 0.386 * 0.35198625 0.41946125
## 174 list1 align1_rep2 74 0.373 * 0.34134625 0.41104375
## 175 list1 align1_rep2 75 0.352 * 0.32153250 0.37702250
## 176 list1 align1_rep2 76 0.182 * 0.16085375 0.21122625
## 177 list1 align1_rep2 77 0.200 * 0.18640500 0.22994500
## 178 list1 align1_rep2 78 0.282 * 0.25094500 0.31284375
## 179 list1 align1_rep2 79 0.196 * 0.17596500 0.21995000
## 180 list1 align1_rep2 80 0.184 * 0.16398750 0.21229750
## 181 list1 align1_rep2 81 0.271 * 0.24515250 0.30618125
## 182 list1 align1_rep2 82 0.261 * 0.23753500 0.28403625
## 183 list1 align1_rep2 83 0.308 * 0.28437625 0.34414250
## 184 list1 align1_rep2 84 0.314 * 0.27973375 0.34232875
## 185 list1 align1_rep2 85 0.225 * 0.19278625 0.25966375
## 186 list1 align1_rep2 86 0.190 * 0.16850750 0.22585125
## 187 list1 align1_rep2 87 0.290 * 0.25720375 0.31779125
## 188 list1 align1_rep2 88 0.375 * 0.34532000 0.41309250
## 189 list1 align1_rep2 89 0.263 * 0.22966875 0.29141750
## 190 list1 align1_rep2 90 0.148 * 0.12857000 0.16226250
## 191 list1 align1_rep2 91 0.258 * 0.22630750 0.28459375
## 192 list1 align1_rep2 92 0.267 * 0.23366000 0.29987625
## 193 list1 align1_rep2 93 0.164 * 0.14525750 0.18582625
## 194 list1 align1_rep2 94 0.162 * 0.14489375 0.17667750
## 195 list1 align1_rep2 95 0.133 * 0.10982875 0.15020250
## 196 list1 align1_rep2 96 0.098 * 0.08089500 0.11271375
## 197 list1 align1_rep2 97 0.117 * 0.09767250 0.14115125
## 198 list1 align1_rep2 98 0.179 * 0.15368875 0.19948875
## 199 list1 align1_rep2 99 0.190 * 0.16731625 0.21410500
## 200 list1 align1_rep2 100 0.185 * 0.15999375 0.20724375
## 201 list1 align2_rep1 1 0.087 * 0.07171875 0.10266750
## 202 list1 align2_rep1 2 0.148 * 0.12442250 0.17715125
## 203 list1 align2_rep1 3 0.175 * 0.14818750 0.19446500
## 204 list1 align2_rep1 4 0.125 * 0.11059875 0.14079750
## 205 list1 align2_rep1 5 0.097 * 0.07764875 0.11480375
## 206 list1 align2_rep1 6 0.060 * 0.04962500 0.06742000
## 207 list1 align2_rep1 7 0.130 * 0.10995125 0.15419250
## 208 list1 align2_rep1 8 0.199 * 0.17599000 0.22295375
## 209 list1 align2_rep1 9 0.166 * 0.14601125 0.18805375
## 210 list1 align2_rep1 10 0.104 * 0.08754250 0.11799875
## 211 list1 align2_rep1 11 0.049 * 0.03787125 0.06093250
## 212 list1 align2_rep1 12 0.031 * 0.02466125 0.03464750
## 213 list1 align2_rep1 13 0.117 * 0.09714125 0.13447125
## 214 list1 align2_rep1 14 0.170 * 0.14637375 0.18907125
## 215 list1 align2_rep1 15 0.163 * 0.14328875 0.19373875
## 216 list1 align2_rep1 16 0.127 * 0.10920375 0.15143625
## 217 list1 align2_rep1 17 0.123 * 0.10245000 0.14502250
## 218 list1 align2_rep1 18 0.183 * 0.16563375 0.20110000
## 219 list1 align2_rep1 19 0.201 * 0.17657375 0.22260875
## 220 list1 align2_rep1 20 0.146 * 0.13190250 0.16459125
## 221 list1 align2_rep1 21 0.153 * 0.13469375 0.16890500
## 222 list1 align2_rep1 22 0.169 * 0.14025500 0.19808375
## 223 list1 align2_rep1 23 0.163 * 0.13066750 0.19421375
## 224 list1 align2_rep1 24 0.151 * 0.12723375 0.17435875
## 225 list1 align2_rep1 25 0.222 * 0.19421750 0.25058875
## 226 list1 align2_rep1 26 0.241 * 0.21796750 0.26165250
## 227 list1 align2_rep1 27 0.230 * 0.20746250 0.25428375
## 228 list1 align2_rep1 28 0.249 * 0.22345250 0.27210375
## 229 list1 align2_rep1 29 0.280 * 0.24924375 0.31301125
## 230 list1 align2_rep1 30 0.255 * 0.23061000 0.27587125
## 231 list1 align2_rep1 31 0.173 * 0.15447375 0.19604875
## 232 list1 align2_rep1 32 0.164 * 0.14018000 0.17966375
## 233 list1 align2_rep1 33 0.275 * 0.24689875 0.30736000
## 234 list1 align2_rep1 34 0.247 * 0.22012875 0.27713000
## 235 list1 align2_rep1 35 0.245 * 0.21815375 0.26779000
## 236 list1 align2_rep1 36 0.299 * 0.27448875 0.32973375
## 237 list1 align2_rep1 37 0.337 * 0.30910625 0.36200375
## 238 list1 align2_rep1 38 0.313 * 0.27748000 0.34524625
## 239 list1 align2_rep1 39 0.257 * 0.23209375 0.28312000
## 240 list1 align2_rep1 40 0.252 * 0.22317375 0.27516000
## 241 list1 align2_rep1 41 0.305 * 0.27300875 0.32798250
## 242 list1 align2_rep1 42 0.371 * 0.31522250 0.41935500
## 243 list1 align2_rep1 43 0.332 * 0.29944750 0.36717375
## 244 list1 align2_rep1 44 0.345 * 0.30523250 0.37523750
## 245 list1 align2_rep1 45 0.318 * 0.29453875 0.34396125
## 246 list1 align2_rep1 46 0.282 * 0.26434750 0.30304625
## 247 list1 align2_rep1 47 0.302 * 0.27026125 0.33677500
## 248 list1 align2_rep1 48 0.374 * 0.35126250 0.39772625
## 249 list1 align2_rep1 49 0.306 * 0.28384625 0.33743625
## 250 list1 align2_rep1 50 0.411 * 0.37962000 0.43936500
## 251 list1 align2_rep1 51 0.649 * 0.59616875 0.70362250
## 252 list1 align2_rep1 52 0.614 * 0.57242375 0.65796000
## 253 list1 align2_rep1 53 0.476 * 0.44010000 0.50812125
## 254 list1 align2_rep1 54 0.430 * 0.39647250 0.45777625
## 255 list1 align2_rep1 55 0.436 * 0.40130000 0.46207625
## 256 list1 align2_rep1 56 0.426 * 0.39785250 0.46162750
## 257 list1 align2_rep1 57 0.396 * 0.36297375 0.42139500
## 258 list1 align2_rep1 58 0.395 * 0.35813250 0.42211375
## 259 list1 align2_rep1 59 0.379 * 0.31963625 0.41232125
## 260 list1 align2_rep1 60 0.304 * 0.27353500 0.33064875
## 261 list1 align2_rep1 61 0.283 * 0.25941000 0.30965500
## 262 list1 align2_rep1 62 0.371 * 0.33135125 0.40681250
## 263 list1 align2_rep1 63 0.286 * 0.26098500 0.31256250
## 264 list1 align2_rep1 64 0.211 * 0.18426500 0.22980375
## 265 list1 align2_rep1 65 0.259 * 0.21719625 0.28183750
## 266 list1 align2_rep1 66 0.258 * 0.23199000 0.29132500
## 267 list1 align2_rep1 67 0.252 * 0.21706750 0.27661000
## 268 list1 align2_rep1 68 0.200 * 0.17929000 0.22251500
## 269 list1 align2_rep1 69 0.140 * 0.12221375 0.16450000
## 270 list1 align2_rep1 70 0.092 * 0.08069000 0.11146750
## 271 list1 align2_rep1 71 0.140 * 0.12411375 0.15536500
## 272 list1 align2_rep1 72 0.238 * 0.20361750 0.26848500
## 273 list1 align2_rep1 73 0.255 * 0.21756875 0.28500375
## 274 list1 align2_rep1 74 0.266 * 0.22368000 0.30308875
## 275 list1 align2_rep1 75 0.256 * 0.22748125 0.27800250
## 276 list1 align2_rep1 76 0.222 * 0.20252375 0.24986875
## 277 list1 align2_rep1 77 0.157 * 0.13760125 0.17630375
## 278 list1 align2_rep1 78 0.199 * 0.17974125 0.22483750
## 279 list1 align2_rep1 79 0.294 * 0.26329500 0.32791375
## 280 list1 align2_rep1 80 0.232 * 0.20800750 0.25773000
## 281 list1 align2_rep1 81 0.177 * 0.15445250 0.20417375
## 282 list1 align2_rep1 82 0.158 * 0.13989625 0.17520875
## 283 list1 align2_rep1 83 0.166 * 0.14832125 0.18908625
## 284 list1 align2_rep1 84 0.168 * 0.14990750 0.18705875
## 285 list1 align2_rep1 85 0.247 * 0.22016000 0.27200250
## 286 list1 align2_rep1 86 0.276 * 0.24482875 0.30513750
## 287 list1 align2_rep1 87 0.162 * 0.14482000 0.17572750
## 288 list1 align2_rep1 88 0.064 * 0.05300625 0.07536500
## 289 list1 align2_rep1 89 0.056 * 0.04384125 0.07048125
## 290 list1 align2_rep1 90 0.129 * 0.11004000 0.15092875
## 291 list1 align2_rep1 91 0.191 * 0.16600250 0.21677250
## 292 list1 align2_rep1 92 0.199 * 0.16086250 0.23012750
## 293 list1 align2_rep1 93 0.167 * 0.15168375 0.18805500
## 294 list1 align2_rep1 94 0.150 * 0.13577250 0.17013375
## 295 list1 align2_rep1 95 0.127 * 0.11447375 0.14289250
## 296 list1 align2_rep1 96 0.143 * 0.12637875 0.16358125
## 297 list1 align2_rep1 97 0.130 * 0.11084125 0.15194625
## 298 list1 align2_rep1 98 0.118 * 0.09970000 0.13721500
## 299 list1 align2_rep1 99 0.116 * 0.09143375 0.13809125
## 300 list1 align2_rep1 100 0.191 * 0.16923500 0.21676625
## 301 list1 align2_rep2 1 0.150 * 0.11281625 0.17883500
## 302 list1 align2_rep2 2 0.132 * 0.09932000 0.16737500
## 303 list1 align2_rep2 3 0.139 * 0.10925750 0.17505750
## 304 list1 align2_rep2 4 0.320 * 0.24536000 0.41013000
## 305 list1 align2_rep2 5 0.677 * 0.54397000 0.77122875
## 306 list1 align2_rep2 6 0.463 * 0.37020375 0.56800625
## 307 list1 align2_rep2 7 0.033 * 0.02431875 0.04350750
## 308 list1 align2_rep2 8 0.080 * 0.05390000 0.10277500
## 309 list1 align2_rep2 9 0.048 * 0.03045125 0.06176875
## 310 list1 align2_rep2 10 0.020 * 0.01300000 0.03110000
## 311 list1 align2_rep2 11 0.015 * 0.01102875 0.01778875
## 312 list1 align2_rep2 12 0.021 * 0.01390125 0.03093000
## 313 list1 align2_rep2 13 0.029 * 0.02092875 0.03715875
## 314 list1 align2_rep2 14 0.153 * 0.12016500 0.18822125
## 315 list1 align2_rep2 15 0.220 * 0.16045000 0.26765000
## 316 list1 align2_rep2 16 0.177 * 0.14525625 0.21320000
## 317 list1 align2_rep2 17 0.342 * 0.28179250 0.40074500
## 318 list1 align2_rep2 18 0.435 * 0.30511250 0.50721625
## 319 list1 align2_rep2 19 0.603 * 0.47183000 0.80414125
## 320 list1 align2_rep2 20 0.400 * 0.29114250 0.48734000
## 321 list1 align2_rep2 21 0.277 * 0.23104750 0.32090750
## 322 list1 align2_rep2 22 0.287 * 0.21435750 0.38468125
## 323 list1 align2_rep2 23 0.277 * 0.22784375 0.32614375
## 324 list1 align2_rep2 24 0.480 * 0.42838500 0.54167500
## 325 list1 align2_rep2 25 0.477 * 0.40615500 0.54803125
## 326 list1 align2_rep2 26 0.364 * 0.28097875 0.45413750
## 327 list1 align2_rep2 27 0.373 * 0.29006125 0.44548875
## 328 list1 align2_rep2 28 0.558 * 0.41009000 0.69051250
## 329 list1 align2_rep2 29 0.397 * 0.33392250 0.48211500
## 330 list1 align2_rep2 30 0.266 * 0.20092875 0.32135250
## 331 list1 align2_rep2 31 0.259 * 0.19049750 0.31885625
## 332 list1 align2_rep2 32 0.555 * 0.45820625 0.64537125
## 333 list1 align2_rep2 33 0.247 * 0.20206125 0.29266375
## 334 list1 align2_rep2 34 0.163 * 0.12842125 0.19553875
## 335 list1 align2_rep2 35 0.275 * 0.21078375 0.33100625
## 336 list1 align2_rep2 36 0.155 * 0.13179000 0.18178250
## 337 list1 align2_rep2 37 0.412 * 0.35661875 0.48300500
## 338 list1 align2_rep2 38 0.509 * 0.43617625 0.60264625
## 339 list1 align2_rep2 39 0.128 * 0.10216375 0.14692125
## 340 list1 align2_rep2 40 0.261 * 0.20894875 0.30634250
## 341 list1 align2_rep2 41 0.901 * 0.79348250 0.96665500
## 342 list1 align2_rep2 42 0.811 * 0.70154000 0.90676500
## 343 list1 align2_rep2 43 0.915 * 0.78133375 1.05155375
## 344 list1 align2_rep2 44 1.259 * 1.15791000 1.42934125
## 345 list1 align2_rep2 45 1.376 * 1.25490250 1.49842000
## 346 list1 align2_rep2 46 1.493 * 1.32818375 1.69967750
## 347 list1 align2_rep2 47 1.185 * 1.02520500 1.30768875
## 348 list1 align2_rep2 48 1.324 * 1.20726875 1.48934375
## 349 list1 align2_rep2 49 2.221 * 2.01059250 2.51347125
## 350 list1 align2_rep2 50 2.301 * 2.05538000 2.62544375
## 351 list1 align2_rep2 51 1.610 * 1.46441750 1.79519875
## 352 list1 align2_rep2 52 1.464 * 1.27733750 1.68247625
## 353 list1 align2_rep2 53 1.342 * 1.17164250 1.47077375
## 354 list1 align2_rep2 54 1.217 * 1.11529625 1.30866750
## 355 list1 align2_rep2 55 1.312 * 1.20364000 1.43370250
## 356 list1 align2_rep2 56 1.316 * 1.21369375 1.49522500
## 357 list1 align2_rep2 57 1.322 * 1.12186125 1.53433125
## 358 list1 align2_rep2 58 0.944 * 0.83270750 1.02712375
## 359 list1 align2_rep2 59 0.709 * 0.63177625 0.80052250
## 360 list1 align2_rep2 60 0.642 * 0.51882500 0.72040000
## 361 list1 align2_rep2 61 0.807 * 0.69967125 0.90052125
## 362 list1 align2_rep2 62 0.846 * 0.74153875 0.98842000
## 363 list1 align2_rep2 63 0.693 * 0.62387750 0.77859500
## 364 list1 align2_rep2 64 1.135 * 1.01681375 1.27420125
## 365 list1 align2_rep2 65 0.830 * 0.74218500 0.94950250
## 366 list1 align2_rep2 66 0.297 * 0.24447625 0.34591500
## 367 list1 align2_rep2 67 0.370 * 0.33365750 0.42596250
## 368 list1 align2_rep2 68 0.341 * 0.27244500 0.39222625
## 369 list1 align2_rep2 69 0.306 * 0.23530500 0.39898500
## 370 list1 align2_rep2 70 0.371 * 0.30136625 0.47234750
## 371 list1 align2_rep2 71 0.412 * 0.35071000 0.45839875
## 372 list1 align2_rep2 72 0.378 * 0.32665500 0.43643500
## 373 list1 align2_rep2 73 0.346 * 0.26752375 0.42469625
## 374 list1 align2_rep2 74 0.292 * 0.22277625 0.35682125
## 375 list1 align2_rep2 75 0.140 * 0.11597875 0.16807250
## 376 list1 align2_rep2 76 0.069 * 0.03605250 0.10005000
## 377 list1 align2_rep2 77 0.105 * 0.07027500 0.14325000
## 378 list1 align2_rep2 78 0.137 * 0.09146500 0.17603250
## 379 list1 align2_rep2 79 0.137 * 0.11318125 0.16009625
## 380 list1 align2_rep2 80 0.472 * 0.40214875 0.55493000
## 381 list1 align2_rep2 81 0.456 * 0.35170125 0.54860500
## 382 list1 align2_rep2 82 0.157 * 0.12238000 0.19158500
## 383 list1 align2_rep2 83 0.046 * 0.03211250 0.06473500
## 384 list1 align2_rep2 84 0.151 * 0.11367000 0.19621250
## 385 list1 align2_rep2 85 0.193 * 0.14745625 0.22704750
## 386 list1 align2_rep2 86 0.220 * 0.16716750 0.25555250
## 387 list1 align2_rep2 87 0.499 * 0.36797125 0.60023250
## 388 list1 align2_rep2 88 0.337 * 0.24731750 0.43525875
## 389 list1 align2_rep2 89 0.140 * 0.10365750 0.16587500
## 390 list1 align2_rep2 90 0.203 * 0.17175875 0.24899750
## 391 list1 align2_rep2 91 0.378 * 0.30961125 0.48894375
## 392 list1 align2_rep2 92 0.556 * 0.46054500 0.65569875
## 393 list1 align2_rep2 93 0.284 * 0.22541250 0.35649750
## 394 list1 align2_rep2 94 0.081 * 0.05927250 0.10832625
## 395 list1 align2_rep2 95 0.138 * 0.09860250 0.17849250
## 396 list1 align2_rep2 96 0.361 * 0.28522625 0.44726125
## 397 list1 align2_rep2 97 0.298 * 0.23667000 0.35768250
## 398 list1 align2_rep2 98 0.606 * 0.47812250 0.72915750
## 399 list1 align2_rep2 99 0.536 * 0.43665000 0.63952500
## 400 list1 align2_rep2 100 0.174 * 0.13497000 0.23667750
## 401 list1 ctrl 1 0.584 * 0.52146625 0.63194375
## 402 list1 ctrl 2 0.515 * 0.48106000 0.55689125
## 403 list1 ctrl 3 0.592 * 0.55048250 0.63196875
## 404 list1 ctrl 4 0.654 * 0.59906750 0.71408625
## 405 list1 ctrl 5 0.651 * 0.60301000 0.71351750
## 406 list1 ctrl 6 0.660 * 0.60845875 0.71488500
## 407 list1 ctrl 7 0.614 * 0.58044500 0.64473125
## 408 list1 ctrl 8 0.574 * 0.53825000 0.62075125
## 409 list1 ctrl 9 0.567 * 0.52714500 0.61325500
## 410 list1 ctrl 10 0.568 * 0.51442750 0.60552750
## 411 list1 ctrl 11 0.644 * 0.58271750 0.68079750
## 412 list1 ctrl 12 0.504 * 0.46957875 0.54730125
## 413 list1 ctrl 13 0.487 * 0.43470000 0.53521875
## 414 list1 ctrl 14 0.530 * 0.48795375 0.56390750
## 415 list1 ctrl 15 0.620 * 0.56754750 0.67204500
## 416 list1 ctrl 16 0.708 * 0.66041750 0.76109125
## 417 list1 ctrl 17 0.798 * 0.76135125 0.83922750
## 418 list1 ctrl 18 0.796 * 0.75865500 0.83702000
## 419 list1 ctrl 19 0.855 * 0.79589750 0.90069125
## 420 list1 ctrl 20 0.790 * 0.72293375 0.84434875
## 421 list1 ctrl 21 0.792 * 0.73522375 0.84924250
## 422 list1 ctrl 22 0.854 * 0.78381750 0.92673250
## 423 list1 ctrl 23 0.786 * 0.73146500 0.85204875
## 424 list1 ctrl 24 0.984 * 0.91954500 1.05298250
## 425 list1 ctrl 25 1.143 * 1.07145375 1.23061625
## 426 list1 ctrl 26 0.919 * 0.84838375 1.01978375
## 427 list1 ctrl 27 0.940 * 0.86651875 1.04753500
## 428 list1 ctrl 28 1.023 * 0.94480875 1.10614625
## 429 list1 ctrl 29 1.112 * 1.03091500 1.19110875
## 430 list1 ctrl 30 1.076 * 1.02257750 1.13058375
## 431 list1 ctrl 31 1.020 * 0.95530500 1.08302375
## 432 list1 ctrl 32 1.217 * 1.13937750 1.29795500
## 433 list1 ctrl 33 1.373 * 1.31334500 1.43829875
## 434 list1 ctrl 34 1.663 * 1.59542500 1.71668750
## 435 list1 ctrl 35 1.750 * 1.61531125 1.84679625
## 436 list1 ctrl 36 1.727 * 1.62843250 1.81305250
## 437 list1 ctrl 37 1.786 * 1.69671125 1.88139000
## 438 list1 ctrl 38 2.013 * 1.87940500 2.12335250
## 439 list1 ctrl 39 2.331 * 2.19927125 2.50645250
## 440 list1 ctrl 40 2.336 * 2.15459500 2.46104000
## 441 list1 ctrl 41 2.092 * 1.98675000 2.21901375
## 442 list1 ctrl 42 2.298 * 2.21585375 2.40531875
## 443 list1 ctrl 43 2.821 * 2.67850250 2.97730500
## 444 list1 ctrl 44 3.097 * 2.97402875 3.22243750
## 445 list1 ctrl 45 3.621 * 3.45436625 3.79990000
## 446 list1 ctrl 46 4.533 * 4.34518875 4.76825375
## 447 list1 ctrl 47 4.631 * 4.41542250 4.82358375
## 448 list1 ctrl 48 4.739 * 4.53074500 4.95318125
## 449 list1 ctrl 49 4.906 * 4.76000500 5.09486875
## 450 list1 ctrl 50 5.254 * 5.01246750 5.45337000
## 451 list1 ctrl 51 5.477 * 5.21559375 5.65580125
## 452 list1 ctrl 52 4.813 * 4.60679250 4.98480250
## 453 list1 ctrl 53 4.509 * 4.34115625 4.68986750
## 454 list1 ctrl 54 3.935 * 3.75470000 4.17431375
## 455 list1 ctrl 55 3.140 * 2.96429875 3.32273375
## 456 list1 ctrl 56 2.912 * 2.76422875 3.03727000
## 457 list1 ctrl 57 2.945 * 2.80720250 3.08985875
## 458 list1 ctrl 58 2.748 * 2.63748250 2.89433625
## 459 list1 ctrl 59 2.555 * 2.45354125 2.65031625
## 460 list1 ctrl 60 2.708 * 2.55231750 2.86026625
## 461 list1 ctrl 61 2.500 * 2.41158000 2.64673500
## 462 list1 ctrl 62 2.279 * 2.16083625 2.37464375
## 463 list1 ctrl 63 2.672 * 2.54233375 2.80343000
## 464 list1 ctrl 64 2.878 * 2.72489875 3.01186500
## 465 list1 ctrl 65 2.269 * 2.17030125 2.44191375
## 466 list1 ctrl 66 1.775 * 1.67372000 1.89280500
## 467 list1 ctrl 67 1.684 * 1.56626875 1.77361625
## 468 list1 ctrl 68 1.558 * 1.46707375 1.64350875
## 469 list1 ctrl 69 1.326 * 1.25399500 1.42397625
## 470 list1 ctrl 70 1.319 * 1.23292000 1.43814250
## 471 list1 ctrl 71 1.376 * 1.26594625 1.46212750
## 472 list1 ctrl 72 1.121 * 1.03752000 1.18316250
## 473 list1 ctrl 73 1.067 * 1.01229250 1.14061875
## 474 list1 ctrl 74 1.111 * 1.06086875 1.17542750
## 475 list1 ctrl 75 1.069 * 1.00803250 1.10735000
## 476 list1 ctrl 76 1.086 * 1.00607875 1.14995250
## 477 list1 ctrl 77 0.938 * 0.86726125 1.01388750
## 478 list1 ctrl 78 0.836 * 0.77480500 0.88776875
## 479 list1 ctrl 79 0.831 * 0.77215375 0.88279500
## 480 list1 ctrl 80 0.868 * 0.81705000 0.93527750
## 481 list1 ctrl 81 0.918 * 0.84147250 0.99688875
## 482 list1 ctrl 82 0.690 * 0.64129000 0.74988500
## 483 list1 ctrl 83 0.764 * 0.71242500 0.80697750
## 484 list1 ctrl 84 0.957 * 0.88850625 1.02667750
## 485 list1 ctrl 85 0.839 * 0.77731125 0.87384250
## 486 list1 ctrl 86 0.984 * 0.91469250 1.06100875
## 487 list1 ctrl 87 1.081 * 1.01555125 1.15459000
## 488 list1 ctrl 88 0.868 * 0.80056125 0.93652625
## 489 list1 ctrl 89 0.779 * 0.72101875 0.83599250
## 490 list1 ctrl 90 1.013 * 0.94595125 1.05816625
## 491 list1 ctrl 91 0.984 * 0.92250875 1.06396750
## 492 list1 ctrl 92 0.995 * 0.92303250 1.07072750
## 493 list1 ctrl 93 1.035 * 0.96419125 1.11723000
## 494 list1 ctrl 94 0.810 * 0.73906125 0.88075375
## 495 list1 ctrl 95 0.902 * 0.83931000 0.97056250
## 496 list1 ctrl 96 1.034 * 0.95035750 1.14662500
## 497 list1 ctrl 97 0.903 * 0.78675875 0.97838250
## 498 list1 ctrl 98 0.808 * 0.73436875 0.88826375
## 499 list1 ctrl 99 0.876 * 0.81160625 0.95731875
## 500 list1 ctrl 100 0.655 * 0.56205625 0.71195500
## 501 list2 align1_rep1 1 0.525 * 0.47729375 0.58599500
## 502 list2 align1_rep1 2 0.495 * 0.42828125 0.57111000
## 503 list2 align1_rep1 3 0.469 * 0.42006000 0.51933125
## 504 list2 align1_rep1 4 0.442 * 0.39606625 0.50267625
## 505 list2 align1_rep1 5 0.482 * 0.37314125 0.59188500
## 506 list2 align1_rep1 6 0.627 * 0.52719000 0.77096500
## 507 list2 align1_rep1 7 0.719 * 0.54054500 0.90196250
## 508 list2 align1_rep1 8 0.704 * 0.58591750 0.87379375
## 509 list2 align1_rep1 9 0.777 * 0.66569750 0.96965625
## 510 list2 align1_rep1 10 0.825 * 0.68310875 0.97844125
## 511 list2 align1_rep1 11 0.668 * 0.59310375 0.75953250
## 512 list2 align1_rep1 12 0.564 * 0.48052750 0.67719875
## 513 list2 align1_rep1 13 0.497 * 0.41659000 0.56934375
## 514 list2 align1_rep1 14 0.580 * 0.47922750 0.68747625
## 515 list2 align1_rep1 15 0.652 * 0.55274500 0.72583750
## 516 list2 align1_rep1 16 0.419 * 0.36361750 0.46608500
## 517 list2 align1_rep1 17 0.443 * 0.37833875 0.49217750
## 518 list2 align1_rep1 18 0.517 * 0.44972000 0.59696625
## 519 list2 align1_rep1 19 0.437 * 0.37855250 0.49270000
## 520 list2 align1_rep1 20 0.500 * 0.45883875 0.54059750
## 521 list2 align1_rep1 21 0.702 * 0.64120250 0.75543750
## 522 list2 align1_rep1 22 0.666 * 0.61633000 0.71586000
## 523 list2 align1_rep1 23 0.572 * 0.51232125 0.61545250
## 524 list2 align1_rep1 24 0.594 * 0.53727625 0.66499375
## 525 list2 align1_rep1 25 0.507 * 0.46168750 0.54756375
## 526 list2 align1_rep1 26 0.497 * 0.46154875 0.53641125
## 527 list2 align1_rep1 27 0.602 * 0.54699625 0.66844875
## 528 list2 align1_rep1 28 0.518 * 0.48805875 0.54834500
## 529 list2 align1_rep1 29 0.353 * 0.30881125 0.39514000
## 530 list2 align1_rep1 30 0.329 * 0.29010375 0.36377500
## 531 list2 align1_rep1 31 0.398 * 0.36167875 0.43130000
## 532 list2 align1_rep1 32 0.426 * 0.39600125 0.45490500
## 533 list2 align1_rep1 33 0.404 * 0.38042625 0.43072750
## 534 list2 align1_rep1 34 0.448 * 0.40652000 0.47659750
## 535 list2 align1_rep1 35 0.622 * 0.57762000 0.69463375
## 536 list2 align1_rep1 36 0.621 * 0.54658000 0.68086375
## 537 list2 align1_rep1 37 0.686 * 0.61127625 0.77090625
## 538 list2 align1_rep1 38 0.706 * 0.64032500 0.76041875
## 539 list2 align1_rep1 39 0.596 * 0.53136000 0.65254500
## 540 list2 align1_rep1 40 0.655 * 0.59366000 0.73084625
## 541 list2 align1_rep1 41 0.687 * 0.61632750 0.74323125
## 542 list2 align1_rep1 42 0.784 * 0.72241500 0.85255500
## 543 list2 align1_rep1 43 0.687 * 0.64197000 0.74935125
## 544 list2 align1_rep1 44 0.623 * 0.55851500 0.70296250
## 545 list2 align1_rep1 45 0.844 * 0.76839125 0.89988250
## 546 list2 align1_rep1 46 0.989 * 0.90530625 1.06043500
## 547 list2 align1_rep1 47 1.054 * 0.97544875 1.14645000
## 548 list2 align1_rep1 48 1.022 * 0.95138250 1.11786750
## 549 list2 align1_rep1 49 1.129 * 0.99182250 1.27952375
## 550 list2 align1_rep1 50 1.097 * 0.96843875 1.22815500
## 551 list2 align1_rep1 51 0.865 * 0.79318625 0.91338875
## 552 list2 align1_rep1 52 0.872 * 0.81610875 0.94299250
## 553 list2 align1_rep1 53 0.926 * 0.85811625 0.98120625
## 554 list2 align1_rep1 54 0.811 * 0.76368000 0.87811375
## 555 list2 align1_rep1 55 0.660 * 0.62301000 0.71084125
## 556 list2 align1_rep1 56 0.695 * 0.64746250 0.75135250
## 557 list2 align1_rep1 57 0.696 * 0.64230000 0.73265375
## 558 list2 align1_rep1 58 0.737 * 0.68879375 0.80572000
## 559 list2 align1_rep1 59 0.593 * 0.55693250 0.63952500
## 560 list2 align1_rep1 60 0.756 * 0.67192875 0.84300625
## 561 list2 align1_rep1 61 0.943 * 0.84124500 1.06340875
## 562 list2 align1_rep1 62 0.850 * 0.79252500 0.91852750
## 563 list2 align1_rep1 63 0.826 * 0.74902625 0.94493875
## 564 list2 align1_rep1 64 0.568 * 0.51141250 0.62950500
## 565 list2 align1_rep1 65 0.590 * 0.54134250 0.65729250
## 566 list2 align1_rep1 66 0.691 * 0.60675500 0.76173750
## 567 list2 align1_rep1 67 0.668 * 0.61229125 0.70821750
## 568 list2 align1_rep1 68 0.566 * 0.49209000 0.60950750
## 569 list2 align1_rep1 69 0.512 * 0.48429375 0.55172500
## 570 list2 align1_rep1 70 0.630 * 0.59503750 0.67467125
## 571 list2 align1_rep1 71 0.720 * 0.66312750 0.76785000
## 572 list2 align1_rep1 72 0.652 * 0.58901250 0.69345500
## 573 list2 align1_rep1 73 0.460 * 0.42492750 0.50694250
## 574 list2 align1_rep1 74 0.388 * 0.35430125 0.43212750
## 575 list2 align1_rep1 75 0.589 * 0.51782875 0.65430875
## 576 list2 align1_rep1 76 0.691 * 0.60107875 0.74194875
## 577 list2 align1_rep1 77 0.645 * 0.60129375 0.69095375
## 578 list2 align1_rep1 78 0.520 * 0.48360625 0.55446750
## 579 list2 align1_rep1 79 0.590 * 0.53350875 0.64460250
## 580 list2 align1_rep1 80 0.609 * 0.56673125 0.66396000
## 581 list2 align1_rep1 81 0.530 * 0.47415375 0.58056500
## 582 list2 align1_rep1 82 0.513 * 0.46856750 0.55661125
## 583 list2 align1_rep1 83 0.491 * 0.44877750 0.54150125
## 584 list2 align1_rep1 84 0.433 * 0.39401375 0.49487125
## 585 list2 align1_rep1 85 0.399 * 0.35455250 0.44169375
## 586 list2 align1_rep1 86 0.394 * 0.35598625 0.43578000
## 587 list2 align1_rep1 87 0.454 * 0.42032875 0.49765250
## 588 list2 align1_rep1 88 0.499 * 0.45269500 0.55427000
## 589 list2 align1_rep1 89 0.541 * 0.46618625 0.59802750
## 590 list2 align1_rep1 90 0.465 * 0.41524625 0.52065250
## 591 list2 align1_rep1 91 0.518 * 0.46171875 0.57682500
## 592 list2 align1_rep1 92 0.433 * 0.39900375 0.46160375
## 593 list2 align1_rep1 93 0.477 * 0.43254000 0.52291750
## 594 list2 align1_rep1 94 0.479 * 0.41670250 0.52973125
## 595 list2 align1_rep1 95 0.598 * 0.48035750 0.67329750
## 596 list2 align1_rep1 96 0.736 * 0.62353000 0.83823500
## 597 list2 align1_rep1 97 0.519 * 0.43940625 0.57567625
## 598 list2 align1_rep1 98 0.637 * 0.55861250 0.72243250
## 599 list2 align1_rep1 99 0.771 * 0.67390375 0.84825000
## 600 list2 align1_rep1 100 0.677 * 0.60153000 0.77370250
## 601 list2 align1_rep2 1 0.062 * 0.04628625 0.07188750
## 602 list2 align1_rep2 2 0.112 * 0.09342500 0.12675750
## 603 list2 align1_rep2 3 0.141 * 0.11641875 0.16685375
## 604 list2 align1_rep2 4 0.112 * 0.09575750 0.13488000
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## 612 list2 align1_rep2 12 0.279 * 0.24133750 0.32979250
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## 616 list2 align1_rep2 16 0.106 * 0.08634500 0.12982125
## 617 list2 align1_rep2 17 0.087 * 0.07801125 0.09954250
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## 619 list2 align1_rep2 19 0.296 * 0.25981250 0.33867125
## 620 list2 align1_rep2 20 0.250 * 0.21390750 0.29575000
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## 625 list2 align1_rep2 25 0.164 * 0.13800250 0.18593375
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## 627 list2 align1_rep2 27 0.176 * 0.14516625 0.20966625
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## 635 list2 align1_rep2 35 0.198 * 0.16371500 0.23323250
## 636 list2 align1_rep2 36 0.211 * 0.17940875 0.23785500
## 637 list2 align1_rep2 37 0.282 * 0.24395000 0.30534875
## 638 list2 align1_rep2 38 0.289 * 0.24860250 0.33141625
## 639 list2 align1_rep2 39 0.306 * 0.26126250 0.33598000
## 640 list2 align1_rep2 40 0.261 * 0.22535625 0.29985250
## 641 list2 align1_rep2 41 0.223 * 0.18621500 0.25792625
## 642 list2 align1_rep2 42 0.426 * 0.38236625 0.46541625
## 643 list2 align1_rep2 43 0.374 * 0.33157375 0.41408375
## 644 list2 align1_rep2 44 0.340 * 0.30401125 0.38064250
## 645 list2 align1_rep2 45 0.486 * 0.42419000 0.53131000
## 646 list2 align1_rep2 46 0.428 * 0.37492125 0.47078000
## 647 list2 align1_rep2 47 0.386 * 0.33482875 0.44631500
## 648 list2 align1_rep2 48 0.398 * 0.34410000 0.44177625
## 649 list2 align1_rep2 49 0.386 * 0.35475250 0.43251375
## 650 list2 align1_rep2 50 0.447 * 0.40039125 0.50131875
## 651 list2 align1_rep2 51 0.417 * 0.36923125 0.45685000
## 652 list2 align1_rep2 52 0.378 * 0.35832875 0.40916875
## 653 list2 align1_rep2 53 0.299 * 0.27385375 0.32737125
## 654 list2 align1_rep2 54 0.310 * 0.28272375 0.35200750
## 655 list2 align1_rep2 55 0.356 * 0.32560625 0.39905625
## 656 list2 align1_rep2 56 0.444 * 0.39034375 0.49147750
## 657 list2 align1_rep2 57 0.367 * 0.32821625 0.40080000
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## 660 list2 align1_rep2 60 0.234 * 0.19506625 0.26454125
## 661 list2 align1_rep2 61 0.317 * 0.27047750 0.37064750
## 662 list2 align1_rep2 62 0.388 * 0.35421125 0.42826250
## 663 list2 align1_rep2 63 0.314 * 0.28652375 0.34462000
## 664 list2 align1_rep2 64 0.289 * 0.25233875 0.32279875
## 665 list2 align1_rep2 65 0.316 * 0.26904625 0.34117000
## 666 list2 align1_rep2 66 0.388 * 0.34626250 0.43892875
## 667 list2 align1_rep2 67 0.361 * 0.33143875 0.39093625
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## 675 list2 align1_rep2 75 0.216 * 0.18325625 0.24842250
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## 677 list2 align1_rep2 77 0.346 * 0.29668750 0.40032875
## 678 list2 align1_rep2 78 0.188 * 0.16719125 0.21765250
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## 683 list2 align1_rep2 83 0.080 * 0.06249500 0.09839000
## 684 list2 align1_rep2 84 0.094 * 0.08375125 0.10873250
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## 686 list2 align1_rep2 86 0.128 * 0.10989125 0.15579875
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## 783 list2 align2_rep1 83 0.129 * 0.11172125 0.14622750
## 784 list2 align2_rep1 84 0.099 * 0.08281625 0.12178250
## 785 list2 align2_rep1 85 0.136 * 0.11030625 0.15965875
## 786 list2 align2_rep1 86 0.130 * 0.11597625 0.15393000
## 787 list2 align2_rep1 87 0.093 * 0.07778125 0.11008500
## 788 list2 align2_rep1 88 0.098 * 0.07815750 0.11800750
## 789 list2 align2_rep1 89 0.073 * 0.05687250 0.09482875
## 790 list2 align2_rep1 90 0.046 * 0.03434500 0.05794000
## 791 list2 align2_rep1 91 0.049 * 0.03443500 0.05789875
## 792 list2 align2_rep1 92 0.046 * 0.03552250 0.06029250
## 793 list2 align2_rep1 93 0.052 * 0.03896500 0.06476000
## 794 list2 align2_rep1 94 0.081 * 0.06680750 0.09251500
## 795 list2 align2_rep1 95 0.070 * 0.05783125 0.08305500
## 796 list2 align2_rep1 96 0.074 * 0.06327250 0.08546625
## 797 list2 align2_rep1 97 0.097 * 0.08052375 0.11900625
## 798 list2 align2_rep1 98 0.078 * 0.06241500 0.09505125
## 799 list2 align2_rep1 99 0.063 * 0.05074875 0.07786750
## 800 list2 align2_rep1 100 0.049 * 0.04149625 0.06289625
## 801 list2 align2_rep2 1 0.170 * 0.13257000 0.20367750
## 802 list2 align2_rep2 2 0.202 * 0.16087500 0.28044250
## 803 list2 align2_rep2 3 0.173 * 0.12842000 0.22153125
## 804 list2 align2_rep2 4 0.187 * 0.15381375 0.23588375
## 805 list2 align2_rep2 5 0.366 * 0.27664625 0.46561750
## 806 list2 align2_rep2 6 0.241 * 0.19438250 0.30991750
## 807 list2 align2_rep2 7 0.146 * 0.10910750 0.18334000
## 808 list2 align2_rep2 8 0.207 * 0.15677750 0.25610875
## 809 list2 align2_rep2 9 0.172 * 0.13424000 0.21121125
## 810 list2 align2_rep2 10 0.279 * 0.20444375 0.33870000
## 811 list2 align2_rep2 11 0.266 * 0.18451500 0.35743000
## 812 list2 align2_rep2 12 0.169 * 0.11947125 0.21082000
## 813 list2 align2_rep2 13 0.180 * 0.13192500 0.23832500
## 814 list2 align2_rep2 14 0.031 * 0.02330125 0.04164625
## 815 list2 align2_rep2 15 0.040 * 0.02645000 0.05755000
## 816 list2 align2_rep2 16 0.014 * 0.00840000 0.02084250
## 817 list2 align2_rep2 17 0.000 * 0.00000000 0.00000000
## 818 list2 align2_rep2 18 0.000 * 0.00000000 0.00000000
## 819 list2 align2_rep2 19 0.000 * 0.00000000 0.00000000
## 820 list2 align2_rep2 20 0.000 * 0.00000000 0.00000000
## 821 list2 align2_rep2 21 0.051 * 0.02862375 0.07140000
## 822 list2 align2_rep2 22 0.142 * 0.10604500 0.17998500
## 823 list2 align2_rep2 23 0.095 * 0.05913750 0.14036250
## 824 list2 align2_rep2 24 0.015 * 0.00783750 0.02175000
## 825 list2 align2_rep2 25 0.060 * 0.03600000 0.08100000
## 826 list2 align2_rep2 26 0.139 * 0.10223000 0.18477000
## 827 list2 align2_rep2 27 0.172 * 0.11960000 0.22245500
## 828 list2 align2_rep2 28 0.025 * 0.01278125 0.03375000
## 829 list2 align2_rep2 29 0.000 * 0.00000000 0.00000000
## 830 list2 align2_rep2 30 0.038 * 0.02280000 0.05130000
## 831 list2 align2_rep2 31 0.082 * 0.05762750 0.10482500
## 832 list2 align2_rep2 32 0.250 * 0.17862500 0.33653750
## 833 list2 align2_rep2 33 0.283 * 0.21700125 0.33430000
## 834 list2 align2_rep2 34 0.235 * 0.18717000 0.27946750
## 835 list2 align2_rep2 35 0.216 * 0.15891000 0.28234000
## 836 list2 align2_rep2 36 0.050 * 0.02706000 0.06762500
## 837 list2 align2_rep2 37 0.099 * 0.06711500 0.12980500
## 838 list2 align2_rep2 38 0.294 * 0.19156500 0.37794000
## 839 list2 align2_rep2 39 0.161 * 0.12496125 0.19996625
## 840 list2 align2_rep2 40 0.227 * 0.14646875 0.30420750
## 841 list2 align2_rep2 41 0.128 * 0.09803500 0.14977000
## 842 list2 align2_rep2 42 0.073 * 0.05227875 0.09330000
## 843 list2 align2_rep2 43 0.179 * 0.11815000 0.23554125
## 844 list2 align2_rep2 44 0.115 * 0.08425875 0.14444250
## 845 list2 align2_rep2 45 0.313 * 0.23905625 0.39007125
## 846 list2 align2_rep2 46 0.186 * 0.13699000 0.23847625
## 847 list2 align2_rep2 47 0.110 * 0.08631250 0.13163000
## 848 list2 align2_rep2 48 0.391 * 0.30036125 0.47017000
## 849 list2 align2_rep2 49 0.303 * 0.26027875 0.35489125
## 850 list2 align2_rep2 50 0.190 * 0.13140000 0.23798750
## 851 list2 align2_rep2 51 0.344 * 0.29473750 0.40107000
## 852 list2 align2_rep2 52 0.372 * 0.29314750 0.47999375
## 853 list2 align2_rep2 53 0.188 * 0.13713500 0.23873625
## 854 list2 align2_rep2 54 0.229 * 0.16759250 0.26096375
## 855 list2 align2_rep2 55 0.290 * 0.23076750 0.34052750
## 856 list2 align2_rep2 56 0.189 * 0.14667750 0.22794375
## 857 list2 align2_rep2 57 0.196 * 0.14301000 0.22857500
## 858 list2 align2_rep2 58 0.323 * 0.22882625 0.44295625
## 859 list2 align2_rep2 59 0.205 * 0.16070625 0.27844375
## 860 list2 align2_rep2 60 0.137 * 0.10558750 0.16502125
## 861 list2 align2_rep2 61 0.275 * 0.21147125 0.34784500
## 862 list2 align2_rep2 62 0.384 * 0.30997750 0.47038750
## 863 list2 align2_rep2 63 0.273 * 0.21641625 0.31430500
## 864 list2 align2_rep2 64 0.159 * 0.12223125 0.20446250
## 865 list2 align2_rep2 65 0.312 * 0.24666125 0.38041375
## 866 list2 align2_rep2 66 0.418 * 0.35401250 0.49461875
## 867 list2 align2_rep2 67 0.248 * 0.18786000 0.31243625
## 868 list2 align2_rep2 68 0.083 * 0.06383750 0.10171750
## 869 list2 align2_rep2 69 0.156 * 0.12951750 0.19401750
## 870 list2 align2_rep2 70 0.151 * 0.12792250 0.17523500
## 871 list2 align2_rep2 71 0.169 * 0.13303750 0.21128625
## 872 list2 align2_rep2 72 0.152 * 0.10707500 0.19079500
## 873 list2 align2_rep2 73 0.025 * 0.01762375 0.03850625
## 874 list2 align2_rep2 74 0.032 * 0.02573500 0.03900000
## 875 list2 align2_rep2 75 0.128 * 0.09607000 0.16675000
## 876 list2 align2_rep2 76 0.296 * 0.23674125 0.35608750
## 877 list2 align2_rep2 77 0.259 * 0.20958875 0.33197500
## 878 list2 align2_rep2 78 0.175 * 0.12775250 0.22706125
## 879 list2 align2_rep2 79 0.394 * 0.32637000 0.47035750
## 880 list2 align2_rep2 80 0.385 * 0.32010625 0.46597375
## 881 list2 align2_rep2 81 0.295 * 0.21211375 0.37269625
## 882 list2 align2_rep2 82 0.478 * 0.36398500 0.57478375
## 883 list2 align2_rep2 83 0.252 * 0.17713500 0.29671250
## 884 list2 align2_rep2 84 0.018 * 0.01280250 0.02520000
## 885 list2 align2_rep2 85 0.130 * 0.07723750 0.17493750
## 886 list2 align2_rep2 86 0.243 * 0.18620000 0.30361125
## 887 list2 align2_rep2 87 0.214 * 0.16766000 0.26988750
## 888 list2 align2_rep2 88 0.030 * 0.02264500 0.03753250
## 889 list2 align2_rep2 89 0.147 * 0.09902500 0.17969250
## 890 list2 align2_rep2 90 0.281 * 0.22892500 0.32715625
## 891 list2 align2_rep2 91 0.251 * 0.20400125 0.31336125
## 892 list2 align2_rep2 92 0.145 * 0.10427250 0.18932375
## 893 list2 align2_rep2 93 0.080 * 0.05690000 0.10800000
## 894 list2 align2_rep2 94 0.024 * 0.01756000 0.02920000
## 895 list2 align2_rep2 95 0.088 * 0.06849000 0.10640000
## 896 list2 align2_rep2 96 0.361 * 0.28096125 0.45533250
## 897 list2 align2_rep2 97 0.586 * 0.42377500 0.69512375
## 898 list2 align2_rep2 98 0.313 * 0.22882625 0.42327375
## 899 list2 align2_rep2 99 0.466 * 0.36268250 0.64480500
## 900 list2 align2_rep2 100 0.415 * 0.28965125 0.57172500
## 901 list2 ctrl 1 0.365 * 0.33292250 0.39868500
## 902 list2 ctrl 2 0.296 * 0.26562625 0.32799125
## 903 list2 ctrl 3 0.311 * 0.27256250 0.35939750
## 904 list2 ctrl 4 0.416 * 0.34961125 0.45595000
## 905 list2 ctrl 5 0.536 * 0.46218750 0.58817875
## 906 list2 ctrl 6 0.357 * 0.31446000 0.39624500
## 907 list2 ctrl 7 0.278 * 0.24570750 0.30751000
## 908 list2 ctrl 8 0.340 * 0.28892875 0.39655875
## 909 list2 ctrl 9 0.403 * 0.33701125 0.46410875
## 910 list2 ctrl 10 0.584 * 0.52771875 0.68847875
## 911 list2 ctrl 11 0.500 * 0.46278000 0.55311250
## 912 list2 ctrl 12 0.448 * 0.40276500 0.50545625
## 913 list2 ctrl 13 0.461 * 0.41385500 0.49906625
## 914 list2 ctrl 14 0.493 * 0.42908625 0.54321125
## 915 list2 ctrl 15 0.430 * 0.38165750 0.48827750
## 916 list2 ctrl 16 0.388 * 0.34559875 0.42446000
## 917 list2 ctrl 17 0.466 * 0.41552750 0.51727750
## 918 list2 ctrl 18 0.387 * 0.34518625 0.43833125
## 919 list2 ctrl 19 0.341 * 0.28772250 0.38959875
## 920 list2 ctrl 20 0.379 * 0.33222750 0.43529125
## 921 list2 ctrl 21 0.340 * 0.30699750 0.37961125
## 922 list2 ctrl 22 0.265 * 0.23199375 0.28956250
## 923 list2 ctrl 23 0.323 * 0.29790250 0.35967375
## 924 list2 ctrl 24 0.492 * 0.45178750 0.54216250
## 925 list2 ctrl 25 0.459 * 0.41762375 0.50806250
## 926 list2 ctrl 26 0.288 * 0.25045250 0.31374250
## 927 list2 ctrl 27 0.358 * 0.31549625 0.40492250
## 928 list2 ctrl 28 0.389 * 0.34658125 0.43359500
## 929 list2 ctrl 29 0.373 * 0.33777625 0.41304375
## 930 list2 ctrl 30 0.374 * 0.32590875 0.40443125
## 931 list2 ctrl 31 0.444 * 0.39315500 0.50002500
## 932 list2 ctrl 32 0.381 * 0.34786750 0.44598125
## 933 list2 ctrl 33 0.481 * 0.43277375 0.52518000
## 934 list2 ctrl 34 0.519 * 0.45851000 0.57846250
## 935 list2 ctrl 35 0.576 * 0.51916625 0.65288000
## 936 list2 ctrl 36 0.589 * 0.51882500 0.63943500
## 937 list2 ctrl 37 0.744 * 0.64795750 0.83779500
## 938 list2 ctrl 38 0.742 * 0.65557750 0.84406875
## 939 list2 ctrl 39 0.729 * 0.66589875 0.80664875
## 940 list2 ctrl 40 0.870 * 0.78112500 0.97816125
## 941 list2 ctrl 41 0.719 * 0.63889125 0.78434875
## 942 list2 ctrl 42 0.616 * 0.52690500 0.68130125
## 943 list2 ctrl 43 0.791 * 0.70180375 0.90482500
## 944 list2 ctrl 44 0.907 * 0.82037750 0.98098875
## 945 list2 ctrl 45 0.984 * 0.89727875 1.04456375
## 946 list2 ctrl 46 1.127 * 1.00568875 1.20811375
## 947 list2 ctrl 47 1.295 * 1.19357000 1.41038625
## 948 list2 ctrl 48 1.275 * 1.18690375 1.38323875
## 949 list2 ctrl 49 1.160 * 1.07113000 1.25702500
## 950 list2 ctrl 50 1.024 * 0.94447625 1.09012250
## 951 list2 ctrl 51 0.964 * 0.86072750 1.07027625
## 952 list2 ctrl 52 0.858 * 0.77586375 0.93895250
## 953 list2 ctrl 53 0.808 * 0.74280875 0.86717000
## 954 list2 ctrl 54 0.888 * 0.80361000 0.95324875
## 955 list2 ctrl 55 0.897 * 0.83590625 0.96317625
## 956 list2 ctrl 56 0.697 * 0.63955250 0.76641250
## 957 list2 ctrl 57 0.753 * 0.66945625 0.84605375
## 958 list2 ctrl 58 0.737 * 0.64505875 0.80281750
## 959 list2 ctrl 59 0.552 * 0.48726375 0.63659750
## 960 list2 ctrl 60 0.520 * 0.45577875 0.58104125
## 961 list2 ctrl 61 0.798 * 0.73782125 0.82928000
## 962 list2 ctrl 62 1.014 * 0.93202375 1.09686625
## 963 list2 ctrl 63 0.752 * 0.69163250 0.81127375
## 964 list2 ctrl 64 0.589 * 0.53972375 0.63299750
## 965 list2 ctrl 65 0.544 * 0.49069500 0.58080500
## 966 list2 ctrl 66 0.483 * 0.44271875 0.53105375
## 967 list2 ctrl 67 0.566 * 0.53198375 0.59331000
## 968 list2 ctrl 68 0.663 * 0.60531000 0.71924250
## 969 list2 ctrl 69 0.521 * 0.47330375 0.57417625
## 970 list2 ctrl 70 0.484 * 0.42175000 0.53490500
## 971 list2 ctrl 71 0.412 * 0.35796875 0.47574250
## 972 list2 ctrl 72 0.385 * 0.34212750 0.42451500
## 973 list2 ctrl 73 0.407 * 0.37753500 0.44120375
## 974 list2 ctrl 74 0.540 * 0.49577125 0.58253000
## 975 list2 ctrl 75 0.574 * 0.51098500 0.62709750
## 976 list2 ctrl 76 0.625 * 0.55763375 0.70587375
## 977 list2 ctrl 77 0.655 * 0.57692250 0.74747375
## 978 list2 ctrl 78 0.558 * 0.50488625 0.61097625
## 979 list2 ctrl 79 0.557 * 0.50914750 0.60250375
## 980 list2 ctrl 80 0.529 * 0.47097500 0.56811375
## 981 list2 ctrl 81 0.586 * 0.52770375 0.65593750
## 982 list2 ctrl 82 0.664 * 0.58496250 0.78142875
## 983 list2 ctrl 83 0.529 * 0.48263625 0.59829125
## 984 list2 ctrl 84 0.324 * 0.28590500 0.36099125
## 985 list2 ctrl 85 0.433 * 0.40051500 0.47160750
## 986 list2 ctrl 86 0.504 * 0.47198750 0.53405750
## 987 list2 ctrl 87 0.347 * 0.31994625 0.38439375
## 988 list2 ctrl 88 0.383 * 0.34056875 0.41685500
## 989 list2 ctrl 89 0.415 * 0.36812500 0.46877000
## 990 list2 ctrl 90 0.349 * 0.31394625 0.37477000
## 991 list2 ctrl 91 0.478 * 0.43933125 0.52377250
## 992 list2 ctrl 92 0.593 * 0.53612750 0.63369000
## 993 list2 ctrl 93 0.384 * 0.35014000 0.42673250
## 994 list2 ctrl 94 0.390 * 0.34980375 0.44492375
## 995 list2 ctrl 95 0.544 * 0.46455625 0.63816000
## 996 list2 ctrl 96 0.513 * 0.39482125 0.61768875
## 997 list2 ctrl 97 0.615 * 0.48426125 0.72427250
## 998 list2 ctrl 98 0.705 * 0.59016000 0.85487750
## 999 list2 ctrl 99 0.708 * 0.54757750 0.84505375
## 1000 list2 ctrl 100 0.652 * 0.58629250 0.75555625
## group
## 1 align1_rep1_list1
## 2 align1_rep1_list1
## 3 align1_rep1_list1
## 4 align1_rep1_list1
## 5 align1_rep1_list1
## 6 align1_rep1_list1
## 7 align1_rep1_list1
## 8 align1_rep1_list1
## 9 align1_rep1_list1
## 10 align1_rep1_list1
## 11 align1_rep1_list1
## 12 align1_rep1_list1
## 13 align1_rep1_list1
## 14 align1_rep1_list1
## 15 align1_rep1_list1
## 16 align1_rep1_list1
## 17 align1_rep1_list1
## 18 align1_rep1_list1
## 19 align1_rep1_list1
## 20 align1_rep1_list1
## 21 align1_rep1_list1
## 22 align1_rep1_list1
## 23 align1_rep1_list1
## 24 align1_rep1_list1
## 25 align1_rep1_list1
## 26 align1_rep1_list1
## 27 align1_rep1_list1
## 28 align1_rep1_list1
## 29 align1_rep1_list1
## 30 align1_rep1_list1
## 31 align1_rep1_list1
## 32 align1_rep1_list1
## 33 align1_rep1_list1
## 34 align1_rep1_list1
## 35 align1_rep1_list1
## 36 align1_rep1_list1
## 37 align1_rep1_list1
## 38 align1_rep1_list1
## 39 align1_rep1_list1
## 40 align1_rep1_list1
## 41 align1_rep1_list1
## 42 align1_rep1_list1
## 43 align1_rep1_list1
## 44 align1_rep1_list1
## 45 align1_rep1_list1
## 46 align1_rep1_list1
## 47 align1_rep1_list1
## 48 align1_rep1_list1
## 49 align1_rep1_list1
## 50 align1_rep1_list1
## 51 align1_rep1_list1
## 52 align1_rep1_list1
## 53 align1_rep1_list1
## 54 align1_rep1_list1
## 55 align1_rep1_list1
## 56 align1_rep1_list1
## 57 align1_rep1_list1
## 58 align1_rep1_list1
## 59 align1_rep1_list1
## 60 align1_rep1_list1
## 61 align1_rep1_list1
## 62 align1_rep1_list1
## 63 align1_rep1_list1
## 64 align1_rep1_list1
## 65 align1_rep1_list1
## 66 align1_rep1_list1
## 67 align1_rep1_list1
## 68 align1_rep1_list1
## 69 align1_rep1_list1
## 70 align1_rep1_list1
## 71 align1_rep1_list1
## 72 align1_rep1_list1
## 73 align1_rep1_list1
## 74 align1_rep1_list1
## 75 align1_rep1_list1
## 76 align1_rep1_list1
## 77 align1_rep1_list1
## 78 align1_rep1_list1
## 79 align1_rep1_list1
## 80 align1_rep1_list1
## 81 align1_rep1_list1
## 82 align1_rep1_list1
## 83 align1_rep1_list1
## 84 align1_rep1_list1
## 85 align1_rep1_list1
## 86 align1_rep1_list1
## 87 align1_rep1_list1
## 88 align1_rep1_list1
## 89 align1_rep1_list1
## 90 align1_rep1_list1
## 91 align1_rep1_list1
## 92 align1_rep1_list1
## 93 align1_rep1_list1
## 94 align1_rep1_list1
## 95 align1_rep1_list1
## 96 align1_rep1_list1
## 97 align1_rep1_list1
## 98 align1_rep1_list1
## 99 align1_rep1_list1
## 100 align1_rep1_list1
## 101 align1_rep2_list1
## 102 align1_rep2_list1
## 103 align1_rep2_list1
## 104 align1_rep2_list1
## 105 align1_rep2_list1
## 106 align1_rep2_list1
## 107 align1_rep2_list1
## 108 align1_rep2_list1
## 109 align1_rep2_list1
## 110 align1_rep2_list1
## 111 align1_rep2_list1
## 112 align1_rep2_list1
## 113 align1_rep2_list1
## 114 align1_rep2_list1
## 115 align1_rep2_list1
## 116 align1_rep2_list1
## 117 align1_rep2_list1
## 118 align1_rep2_list1
## 119 align1_rep2_list1
## 120 align1_rep2_list1
## 121 align1_rep2_list1
## 122 align1_rep2_list1
## 123 align1_rep2_list1
## 124 align1_rep2_list1
## 125 align1_rep2_list1
## 126 align1_rep2_list1
## 127 align1_rep2_list1
## 128 align1_rep2_list1
## 129 align1_rep2_list1
## 130 align1_rep2_list1
## 131 align1_rep2_list1
## 132 align1_rep2_list1
## 133 align1_rep2_list1
## 134 align1_rep2_list1
## 135 align1_rep2_list1
## 136 align1_rep2_list1
## 137 align1_rep2_list1
## 138 align1_rep2_list1
## 139 align1_rep2_list1
## 140 align1_rep2_list1
## 141 align1_rep2_list1
## 142 align1_rep2_list1
## 143 align1_rep2_list1
## 144 align1_rep2_list1
## 145 align1_rep2_list1
## 146 align1_rep2_list1
## 147 align1_rep2_list1
## 148 align1_rep2_list1
## 149 align1_rep2_list1
## 150 align1_rep2_list1
## 151 align1_rep2_list1
## 152 align1_rep2_list1
## 153 align1_rep2_list1
## 154 align1_rep2_list1
## 155 align1_rep2_list1
## 156 align1_rep2_list1
## 157 align1_rep2_list1
## 158 align1_rep2_list1
## 159 align1_rep2_list1
## 160 align1_rep2_list1
## 161 align1_rep2_list1
## 162 align1_rep2_list1
## 163 align1_rep2_list1
## 164 align1_rep2_list1
## 165 align1_rep2_list1
## 166 align1_rep2_list1
## 167 align1_rep2_list1
## 168 align1_rep2_list1
## 169 align1_rep2_list1
## 170 align1_rep2_list1
## 171 align1_rep2_list1
## 172 align1_rep2_list1
## 173 align1_rep2_list1
## 174 align1_rep2_list1
## 175 align1_rep2_list1
## 176 align1_rep2_list1
## 177 align1_rep2_list1
## 178 align1_rep2_list1
## 179 align1_rep2_list1
## 180 align1_rep2_list1
## 181 align1_rep2_list1
## 182 align1_rep2_list1
## 183 align1_rep2_list1
## 184 align1_rep2_list1
## 185 align1_rep2_list1
## 186 align1_rep2_list1
## 187 align1_rep2_list1
## 188 align1_rep2_list1
## 189 align1_rep2_list1
## 190 align1_rep2_list1
## 191 align1_rep2_list1
## 192 align1_rep2_list1
## 193 align1_rep2_list1
## 194 align1_rep2_list1
## 195 align1_rep2_list1
## 196 align1_rep2_list1
## 197 align1_rep2_list1
## 198 align1_rep2_list1
## 199 align1_rep2_list1
## 200 align1_rep2_list1
## 201 align2_rep1_list1
## 202 align2_rep1_list1
## 203 align2_rep1_list1
## 204 align2_rep1_list1
## 205 align2_rep1_list1
## 206 align2_rep1_list1
## 207 align2_rep1_list1
## 208 align2_rep1_list1
## 209 align2_rep1_list1
## 210 align2_rep1_list1
## 211 align2_rep1_list1
## 212 align2_rep1_list1
## 213 align2_rep1_list1
## 214 align2_rep1_list1
## 215 align2_rep1_list1
## 216 align2_rep1_list1
## 217 align2_rep1_list1
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get_params
The various parameters used during the initialization of the metagene
object,
the production of the table and the production of the plot are saved and can
be accessed with the get_params
function:
mg <- get_demo_metagene()
mg$get_params()
## $padding_size
## [1] 0
##
## $verbose
## [1] FALSE
##
## $bam_files
## align1_rep1
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep1.bam"
## align1_rep2
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align1_rep2.bam"
## align2_rep1
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep1.bam"
## align2_rep2
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/align2_rep2.bam"
## ctrl
## "/tmp/RtmpuW2WHq/Rinst444212fd9d9a/metagene/extdata/ctrl.bam"
##
## $force_seqlevels
## [1] FALSE
##
## $flip_regions
## [1] FALSE
##
## $assay
## [1] "chipseq"
##
## $df_needs_update
## [1] TRUE
##
## $df_arguments
## [1] ""
##
## $table_needs_update
## [1] TRUE
get_design
To get the design that was used to produce the last version of the table,
you can use the get_design
function:
mg$produce_table(design = get_demo_design())
## produce data table : ChIP-Seq
## Alternatively, it is also possible to add a design without producing the
## table:
mg$add_design(get_demo_design())
mg$get_design()
## Samples align1 align2
## 1 align1_rep1.bam 1 0
## 2 align1_rep2.bam 1 0
## 3 align2_rep1.bam 0 1
## 4 align2_rep2.bam 0 1
## 5 ctrl.bam 2 2
get_bam_count
To get the number of aligned read in a BAM file, you can use the
get_bam_count
function:
mg$get_bam_count(bam_files[1])
## [1] 4635
get_regions
To get all the regions, you can use the get_regions
function:
mg$get_regions()
## GRangesList object of length 2:
## $list1
## GRanges object with 50 ranges and 0 metadata columns:
## seqnames ranges strand
## <Rle> <IRanges> <Rle>
## [1] chr1 16103663-16105662 *
## [2] chr1 23921318-23923317 *
## [3] chr1 34848977-34850976 *
## [4] chr1 36368182-36370181 *
## [5] chr1 36690488-36692487 *
## ... ... ... ...
## [46] chr1 172081530-172083529 *
## [47] chr1 172081796-172083795 *
## [48] chr1 172147016-172149015 *
## [49] chr1 172205805-172207804 *
## [50] chr1 172260642-172262641 *
##
## ...
## <1 more element>
## -------
## seqinfo: 1 sequence from an unspecified genome; no seqlengths
It is also possible to extract a subset of the regions with the get_regions
function:
mg$get_regions(region_names = c(regions[1]))
## GRangesList object of length 1:
## $list1
## GRanges object with 50 ranges and 0 metadata columns:
## seqnames ranges strand
## <Rle> <IRanges> <Rle>
## [1] chr1 16103663-16105662 *
## [2] chr1 23921318-23923317 *
## [3] chr1 34848977-34850976 *
## [4] chr1 36368182-36370181 *
## [5] chr1 36690488-36692487 *
## ... ... ... ...
## [46] chr1 172081530-172083529 *
## [47] chr1 172081796-172083795 *
## [48] chr1 172147016-172149015 *
## [49] chr1 172205805-172207804 *
## [50] chr1 172260642-172262641 *
##
## -------
## seqinfo: 1 sequence from an unspecified genome; no seqlengths
get_raw_coverages
To get the coverages produced during the initialization of the metagene
object, you can use the get_raw_coverages
function. Please note that to save
space, metagene will only extract the coverages in the regions provided.
coverages <- mg$get_raw_coverages()
coverages[[1]]
## RleList of length 22
## $chr10
## integer-Rle of length 130694993 with 1 run
## Lengths: 130694993
## Values : 0
##
## $chr11
## integer-Rle of length 122082543 with 1 run
## Lengths: 122082543
## Values : 0
##
## $chr12
## integer-Rle of length 120129022 with 1 run
## Lengths: 120129022
## Values : 0
##
## $chr13
## integer-Rle of length 120421639 with 1 run
## Lengths: 120421639
## Values : 0
##
## $chr14
## integer-Rle of length 124902244 with 1 run
## Lengths: 124902244
## Values : 0
##
## ...
## <17 more elements>
length(coverages)
## [1] 5
It is also possible to extract a subset of all the coverages by providing the filenames:
coverages <- mg$get_raw_coverages(filenames = bam_files[1:2])
length(coverages)
## [1] 2
get_normalized_coverages
The get_normalized_coverages
function works exactly like the
get_raw_coverages
function except that it returns the coverages in read per
million aligned (RPM).
Every function of metagene (except for the getters) invisibly return a pointer to itself. This means that the functions can be chained:
rg <- get_demo_regions()
bam <- get_demo_bam_files()
d <- get_demo_design()
title <- "Show chain"
mg <- metagene$new(rg, bam)$produce_table(design = d)$plot(title = title)
## produce data table : ChIP-Seq
## produce data frame : ChIP-Seq
## Plot : ChIP-Seq
To copy a metagene object, you have to use the clone
function:
mg_copy <- mg$clone()
While metagene
try to reduce it’s memory usage, it’s possible to run into
memory limits when working with multiple large datasets (especially when there
is a lot of regions with a large width).
One way to avoid this is to analyse each dataset seperately and then merge just before producing the metagene plot:
mg1 <- metagene$new(bam_files = bam_files, regions = regions[1])
mg1$produce_data_frame()
## produce data table : ChIP-Seq
## produce data frame : ChIP-Seq
mg2 <- metagene$new(bam_files = bam_files, regions = regions[2])
mg2$produce_data_frame()
## produce data table : ChIP-Seq
## produce data frame : ChIP-Seq
Then you can extract the data.frame
s and combine them with rbind
:
df1 <- mg1$get_data_frame()
df2 <- mg2$get_data_frame()
df <- rbind(df1, df2)
Finally, you can use the plot_metagene
function to produce the metagene plot:
p <- plot_metagene(df)
## Plot : ChIP-Seq
p + ggplot2::ggtitle("Managing large datasets")
It is possible to compare two metagene profiles using the permutation_test
function provided with the metagene
package. Please note that the permutation
tests functionality is still in development and is expected to change in future
releases.
The first step is to decide which profiles we want to compare and extract the corresponding tables :
tab <- mg$get_table()
tab0 <- tab[which(tab$region == "list1"),]
tab1 <- tab0[which(tab0$design == "align1"),]
tab2 <- tab0[which(tab0$design == "align2"),]
Then we defined to function to use to compare the two profiles. For this, a
companion package of metagene
named similaRpeak provides
multiple metrics.
For this example, we will prepare a function to calculate the RATIO_NORMALIZED_INTERSECT between two profiles:
library(similaRpeak)
perm_fun <- function(profile1, profile2) {
sim <- similarity(profile1, profile2)
sim[["metrics"]][["RATIO_NORMALIZED_INTERSECT"]]
}
We then compare our two profiles using this metric:
ratio_normalized_intersect <-
perm_fun(tab1[, .(moy=mean(value)), by=bin]$moy,
tab2[, .(moy=mean(value)), by=bin]$moy)
ratio_normalized_intersect
## [1] 0.7387951
To check if this value is significant, we can permute the two tables that were used to produce the profile and calculate their RATIO_NORMALIZED_INTERSECT:
permutation_results <- permutation_test(tab1, tab2, sample_size = 50,
sample_count = 1000, FUN = perm_fun)
Finally, we check how often the calculated value is greater than the results of the permutations:
sum(ratio_normalized_intersect >= permutation_results) /
length(permutation_results)
## [1] 0.022