Comments (7)
This is a standard error when a file is not present. What do you get when you do
ls -l /home/common_all_20180418.vcf
Best
Valentina
from freec.
the file is present in the specified directory
-rw-rw-r--. 1 user user 1593039139 Aug 2 15:08 common_all_20180418.vcf.gz
it's also been bgzipped back to previous
from freec.
apologies, I am switching between 2 online servers and they have slightly different directory trees. there was one directory missing between /home/ and the file. Thanks for pointing that out!
I have a second question, when I don't specify a pileup file in the config as mateFile, and instead I use a BAM file,
config file looks like this:
chrLenFile = /home/HG38/Hg38_chromosomeLengthFile.txt
window = 50000
step = 3000
ploidy = 2
breakPointThreshold = 0.75
chrFiles = /home/HG38/chromosomes/
intercept = 1
outputDir = /home/FREEC-ANALYSIS
sex = XY
breakPointType = 2
coefficientOfVaration = 0.05
[sample]
mateFile = /HOME/WGS.bam
inputFormat = BAM
mateOrientation = FR
[BAF]
makePileup=/home/subdir/common_all_20180418.vcf
SNPfile = /home/subdir/common_all_20180418.vcf
minimalCoveragePerPosition = 0
fastaFile = /home/HG38/hg38.fa
I get an error telling saying
Error: /home/FREEC-ANALYSIS/WGS.bam_NewCaptureRegions.bed is an empty file.
..will use 0 SNPs for calculation of the BAF profile
Is this correct for the first iteration of FREEC whilst it makes the necessary minipileup file, or is something missing from the config file?
To get round this I made my own mpileup file with samtools mpileup, and once overcoming the previous problem, it's now running with no errors as of yet.
best,
Ll.
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Thank you for pointing out the problem. It seems that I did not debug the "minipileup" option for WGS, but only for WES, where a target.bed file is provided with exon coordinates. I will check what is gong on.
UPDATE: it seems that your "/home/HG38/Hg38_chromosomeLengthFile.txt" is in a format that FREEC cannot read in function "createBedFileWithChromosomeLengths()". Can you try to use fa.fai format? and then check that WGS.bam_NewCaptureRegions.bed is not empty?
Also, if you use this "makePileup" option, you .BAM file with reads should contain "chr" prefixes for chromosomes.. Is it the case?
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I will have a look at your .BAM reading issue. However, I would not recommend to use
window = 50000
step = 3000
Instead, just comment or remove these parameters and let FREEC find the best window size for your data.
If you want to keep it, rather use
window = 50000
step = 25000
BTW, parameter coefficientOfVaration = 0.05
to detect the best window size is not used, if you already provide window
.
And I would also let FREEC choose the best parameters for the model:
#intercept = 1
You will also greatly improve the output if you use gemMappabilityFile
to mask low mappability regions.
from freec.
Thank you for pointing out the problem. It seems that I did not debug the "minipileup" option for WGS, but only for WES, where a target.bed file is provided with exon coordinates. I will check what is gong on.
I am running into the same issue outlined above, where the WGS workflow is not properly handling a BAM file without chr prefix in the chromosome names. Has this been fixed in some way that I have missed?
UPDATE: it seems that your "/home/HG38/Hg38_chromosomeLengthFile.txt" is in a format that FREEC cannot read in function "createBedFileWithChromosomeLengths()". Can you try to use fa.fai format? and then check that WGS.bam_NewCaptureRegions.bed is not empty?
Also, if you use this "makePileup" option, you .BAM file with reads should contain "chr" prefixes for chromosomes.. Is it the case?
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v11.5 takes .fai into account in function "createBedFileWithChromosomeLengths()". Is it the version you are using?
Yes, I think that if you create a miniPileup, .BAM files should contain "chr" prefixes.
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