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DepthOfCoverage and DiagnoseTargets: our plans for development and support

We have decided to continue providing and supporting DepthOfCoverage and DiagnoseTargets for the foreseeable future. Going forward, we'll try to integrate them and develop their features to address the main needs of the community. To this end we welcome your continuing feedback, so please feel free to contribute comments and ideas in this thread.

To all who took the time to tell us what you find useful about DoC and DT (and what you wish it could do), a big thank you! This is always very useful to us because it helps us identify which features are most valuable to our users.

Geraldine Van der Auwera, PhD

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• Posts: 2Member

Thanks Geraldine. A newbie question: what's the difference between DepthOfCoverage and DiagnoseTargets, and which one do you recommend to use (or both)?

The two are somewhat convergent, but basically DepthOfCoverage allows you to evaluate the depth of coverage in your data at certain sites or over intervals generally, whereas DiagnoseTargets is more specifically designed to evaluate the quality of data covering exome target intervals. The best thing to do is to read their respective documentations pages (links below) and see which sounds like it will suit you needs best.

Geraldine Van der Auwera, PhD

• Posts: 11Member
edited June 2013

Hi @Geraldine_VdAuwera and thanks for support on this.

The DepthOfCoverage documentation mentions -nt as a parallelism option but I get an error.

##### ERROR MESSAGE: Invalid command line: Argument nt has a bad value: The analysis DepthOfCoverage aggregates results by interval. Due to a current limitation of the GATK, analyses of this type do not currently support parallel execution. Please run your analysis without the -nt option.

My command line:
java -jar GenomeAnalysisTK.jar -T DepthOfCoverage -I ExampleBAM.bam -R exampleFASTA.fasta -o ./test_coverage_out -nt 4

Is this an error in the documentation? Or am I missing something?

Thanks.

Post edited by kevyin on

Hi @kevyin,

DOC is currently set up to aggregate statistics over intervals by default, which is incompatible with the -nt mode. You can disable this behavior by using the --omitIntervalStatistics flag, which should make -nt work. Let me know if you have any issues with that.

We may change the default behavior in future; in any case we will add a note about this to the documentation. Thanks for reporting this!

Geraldine Van der Auwera, PhD

• Posts: 7Member

Hello, I want the concise depth of coverage summary as well as the per-location covering number of bases for my RNA-Seq data.
With not splitting the N containing reads, what does DoC exactly do with -U ALLOW_N_CIGAR_READS option? Is it still counting the bases in the CIGAR array represented as matched? I think the bases represented as N in CIGAR should not be counted, just as the deletions (D) are treated. (Maybe an option such as --includeNBases might be appropriate for special cases.)
A good explanation will be helpful to decide the DoC or not.

And please do something with the last line of the ERROR MESSAGE "Notice however that if you were to choose the latter, an unspecified subset of the analytical outputs of an unspecified subset of the tools will become unpredictable.", which I think is totally nonsense!

Hah, I think the developer who wrote that error message was feeling particularly sarcastic that day. I'll see what we can do to make that a little more informative.

Regarding the behavior question, I believe the Ns in the CIGAR are correctly interpreted as not contributing any coverage.

That said, you could run DoC after running the SplitNTrim step, which will give you effective depth after processing.

Geraldine Van der Auwera, PhD

• Posts: 7Member
• Beijing Institute of Genomics, CASPosts: 51Member

i try to use -nt option while running DoC tool. I added -omitIntervalStatistics. But it still prompts an error.

My command was:
java -Xmx15g -jar /softwares/GTK/GenomeAnalysisTK.jar -T DepthOfCoverage \
-log DepthofCov/depth.log -nt 15 -omitIntervalStatistics \
-R /Ref/human_g1k_v37.fasta \
-I /DepthofCov/CHG000691_2_3bam.list \
-geneList /Ref/humanRefseqROD_Final.refseq \
-o /DepthofCov/HM_trios_Samples

Error Message:

ERROR ------------------------------------------------------------------------------------------

Can you help me how to resolve this issue?
Thanks..