Description Usage Arguments Value Author(s) Examples
This function tests for differential RNA methylation between two conditions.
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control_ip |
a reads count data frame for IP samples of control |
treated_ip |
a reads count data frame for IP samples of treated |
control_input |
a reads count data frame for input samples of control |
treated_input |
a reads count data frame for input samples of treated |
size.factor |
A list of size factor. The size factor of the IP and input sample of the biological replicate and directly reflect their sequencing depth. If size.factor=NA, QNB will compute the size factor of IP and input sample of each replicate. If user could provide the size.factor, the name of each term must be control_ip, treated_ip, control_input, treated_input in list. |
mode |
There are four ways how the empirical dispersion can be computed:
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plot.dispersion |
The default is TRUE. If plot.dispersion=FALSE, it will not save the dispersion figure. |
output.dir |
The saved file path. The default is NA. If output.dir=NA, the path is the current path. |
Results will get a matrix including 7 columns (p.treated, p.control, log2.RR, log2.OR, pvalue, q, padj).
p.treated |
The percentage of methylation under treated condition. |
p.control |
The percentage of methylation under control condition. |
log2.RR |
The normalized risk ratio. |
log2.OR |
The normalized odds ratio. |
pvalue |
Indicate the significance of the methylation site as an RNA differential methylation site |
q |
The standardized feature abundance, which is proportional to the expression level of the RNA transcript. |
padj |
The FDR of the methylation site, indicating the significance of the peak as an RNA differential methylation site after multiple hypothesis correction usting BH method. |
Lian Liu <liulian19860905@163.com>
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 | ## The function is currently defined as load library and specify the parameters
library("QNB")
f1 = system.file("extdata", "control_ip.txt", package="QNB")
f2 = system.file("extdata", "treated_ip.txt", package="QNB")
f3 = system.file("extdata", "control_input.txt", package="QNB")
f4 = system.file("extdata", "treated_input.txt", package="QNB")
meth1 = read.table(f1,header=TRUE)
meth2 = read.table(f2,header=TRUE)
unmeth1 = read.table(f3,header=TRUE)
unmeth2 = read.table(f4,header=TRUE)
# When there are replicates under two conditions, we could select
# mode="per-condition" or mode="pooled" to estimate the dispersion.
# The default is mode="auto".
result = qnbtest(meth1,meth2,unmeth1,unmeth2,mode="per-condition")
# When size.factor is not NA
## Not run:
total_number_reads_control_ip <- c(3015921,2563976,198530)
total_number_reads_treated_ip <- c(1565101,152389,323569)
total_number_reads_control_input <- c(108561,302534,108123)
total_number_reads_treated_input <- c(301270,208549,308654)
# calculate the number of reads for a "standard" library
standard_library_size <- exp(mean(log( c(total_number_reads_control_ip,
total_number_reads_treated_ip,
total_number_reads_control_input,
total_number_reads_treated_input))))
# calculate the sample size factor based on the total number of reads
size.factor <- list(control_ip = total_number_reads_control_ip/standard_library_size,
treated_ip = total_number_reads_treated_ip/standard_library_size,
control_input = total_number_reads_control_input/standard_library_size,
treated_input = total_number_reads_treated_input/standard_library_size)
# use size factor calculated from previous step in QNB model
result <- qnbtest(meth1, meth2, unmeth1, unmeth2,
size.factor = size.factor)
## End(Not run)
# If you have replicates for one condition but not for the other, or without any
# replicates for # two conditions, you can select mode="blind" to estimate
# the dispersion.
f1 = system.file("extdata", "no_rep_controlip.txt", package="QNB")
f2 = system.file("extdata", "no_rep_treatedip.txt", package="QNB")
f3 = system.file("extdata", "no_rep_controlinput.txt", package="QNB")
f4 = system.file("extdata", "no_rep_treatedinput.txt", package="QNB")
no_rep_meth1 = read.table(f1,header=TRUE)
no_rep_meth2 = read.table(f2,header=TRUE)
no_rep_unmeth1 = read.table(f3,header=TRUE)
no_rep_unmeth2 = read.table(f4,header=TRUE)
## Not run:
result = qnbtest(no_rep_meth1,
no_rep_meth2,
no_rep_unmeth1,
no_rep_unmeth2,
mode="blind")
## End(Not run)
# If you could not decide which mode to estimate dispersion, mode="auto"
# will select suitable way to estimate dispersion according to the replicates.
## Not run:
result = qnbtest(meth1, meth2,unmeth1,unmeth2)
## End(Not run)
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