# Tagged with #local realignment 1 documentation article | 0 announcements | 4 forum discussions

Created 2012-07-23 16:48:55 | Updated 2012-09-30 23:35:55 | Tags: indelrealigner realignertargetcreator official

## Realigner Target Creator

For a complete, detailed argument reference, refer to the GATK document page here.

## Indel Realigner

For a complete, detailed argument reference, refer to the GATK document page here.

# Running the Indel Realigner only at known sites

While we advocate for using the Indel Realigner over an aggregated bam using the full Smith-Waterman alignment algorithm, it will work for just a single lane of sequencing data when run in -knownsOnly mode. Novel sites obviously won't be cleaned up, but the majority of a single individual's short indels will already have been seen in dbSNP and/or 1000 Genomes. One would employ the known-only/lane-level realignment strategy in a large-scale project (e.g. 1000 Genomes) where computation time is severely constrained and limited. We modify the example arguments from above to reflect the command-lines necessary for known-only/lane-level cleaning.

The RealignerTargetCreator step would need to be done just once for a single set of indels; so as long as the set of known indels doesn't change, the output.intervals file from below would never need to be recalculated.

 java -Xmx1g -jar /path/to/GenomeAnalysisTK.jar \
-T RealignerTargetCreator \
-R /path/to/reference.fasta \
-o /path/to/output.intervals \
-known /path/to/indel_calls.vcf


The IndelRealigner step needs to be run on every bam file.

java -Xmx4g -Djava.io.tmpdir=/path/to/tmpdir \
-jar /path/to/GenomeAnalysisTK.jar \
-I <lane-level.bam> \
-R <ref.fasta> \
-T IndelRealigner \
-targetIntervals <intervalListFromStep1Above.intervals> \
-o <realignedBam.bam> \
-known /path/to/indel_calls.vcf
--consensusDeterminationModel KNOWNS_ONLY \
-LOD 0.4

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Created 2014-01-31 21:16:16 | Updated | Tags: knownsites dbsnp indel-realignment

Dear GATK team,

Would you please clarify that, based on your experience or the logic used in the realignment algorithm, which option between using dbSNP, 1K gold standard (mills...), or "no known dbase" might result in a more accurate set of indels in the Indel-based realignment stage (speed and efficiency is not my concern).

Based on the documentation I found on your site, the "known" variants are used to identify "intervals" of interest to then perform re-alignment around indels. So, it makes sense to me to use as many number of indels as possible (even if they are unreliable and garbage such as many of those found in dbSNP) in addition to those more accurate calls found in 1K gold-standard datasets for choosing the intervals. After all, that increases he number of indel regions to be investigated and therefore potentially increase the accuracy. Depending on your algorithm logic, also, it seems that providing no known dbase would increase the chance of investigating more candidates of mis-alignment and therefore improving the accuracy.

But if your logic uses the "known" indel sets to just "not" perform the realignment and ignore those candidates around known sites, it makes sense to use the more accurate set such as 1K gold standard.

Please let me know what you suggest.

Thank you Regards Amin Zia

Created 2014-01-31 21:15:22 | Updated | Tags:

Dear GATK team,

Would you please clarify that, based on your experience or the logic used in the realignment algorithm, which option between using dbSNP, 1K gold standard (mills...), or "no known dbase" might result in a more accurate set of indels in the Indel-based realignment stage (speed and efficiency is not my concern).

Based on the documentation I found on your site, the "known" variants are used to identify "intervals" of interest to then perform re-alignment around indels. So, it makes sense to me to use as many number of indels as possible (even if they are unreliable and garbage such as many of those found in dbSNP) in addition to those more accurate calls found in 1K gold-standard datasets for choosing the intervals. After all, that increases he number of indel regions to be investigated and therefore potentially increase the accuracy. Depending on your algorithm logic, also, it seems that providing no known dbase would increase the chance of investigating more candidates of mis-alignment and therefore improving the accuracy.

But if your logic uses the "known" indel sets to just "not" perform the realignment and ignore those candidates around known sites, it makes sense to use the more accurate set such as 1K gold standard.

Please let me know what you suggest.

Thank you Regards Amin Zia

Created 2013-05-22 15:47:54 | Updated | Tags: indelrealigner realignertargetcreator

Hi

I've followed the suggested protocol for local realignment - first using RealignerTargetCreator and then IndelRealigner, but have unexpected results.

Let's call the two BAMs I'm realigning "normal" and "tumour" or N and T for short. Once realigned, I've split the resulting NT BAM file (using readgroup tags, although I see from the docs that it can create separate files natively) back into the original N and T BAM files and discovered something odd. I was expecting the pre-realignment N and T files to contain the same number of reads as the post-realignment files, only the coordinates that reads are mapped to would be different.

However, I notice that post-realignment files contain significantly fewer reads because unaligned reads and reads not aligned to the autosomes or sex chromosomes have been removed. However, these reads alone do not account for the difference; large numbers of reads aligned to the 24 chromosomes are now missing.

Can you tell me more about the reads that are removed? I suspect it to be an alignment quality issue, but cannot find direct reference to this behaviour in the documentation. I'm currently keeping both my pre and post-realignment bam files, but ultimately there will be space constraints and I'll have to choose and would like to make the most informed decision possible.

Regards Chris

Created 2012-07-27 12:12:08 | Updated 2012-07-27 12:12:08 | Tags:

Dear GATK team,

I understood from the best practice documentation that if I have no known indels list for the genome I can skip the realignment step. Is this correct, or shall I call Indels and then do realignment and then re-call SNPs and Indels with the UnifiedGenotyper?

Thanks for the help!