NGS: Indel Analysis tool order
Hello! I have a question about the NGS: Indel Analysis toolset in Galaxy. I have aligned my samples from Illumina's HiSeq2000 to the reference genome using BWA. I've called SNPs using SAMTools and now need to call indels. Under the NGS: Indel Analysis toolset, I see two options: "Filter Indels for SAM" and "Extract indels from SAM." Since the descriptions are a little vague, I would presume that I start with the "Filter Indels for SAM" first on the BWA SAM file and then run "Extract indels from SAM" on the "Filter" output SAM file (i.e. BWA SAM -> Filter Indels for SAM -> Extract indels from SAM). Is this the correct order? Or would I skip the "Filter Indels for SAM" step (since the BWA SAM file technically already contains indels so there would be no need to filter) and just go straight to "Extract indels from SAM" (i.e. BWA SAM -> Extract indels from SAM)? I've tried both ways and get different results. For example: 1. BWA SAM -> Filter Indels for SAM -> Extract indels from SAM -> gives me 26,417 regions of interest 2. BWA SAM -> Extract indels from SAM -> gives me 93,974 regions of interest Which way is correct? Any help/info would be greatly appreciated! Thanks, David
Hello, The "Filter Indels" tool is intended to act as a quality screen before "Extract Indels" is used. Whether you want to use this step is something you will need to determine, as are the exact criteria for filtering. The best advice is to examine a sample of the differences between the two methods and tune or exclude the Filter step. Hopefully this helps, Best, Jen Galaxy team On 8/22/11 11:58 AM, David K Crossman wrote:
Hello!
I have a question about the NGS: Indel Analysis toolset in Galaxy. I have aligned my samples from Illumina’s HiSeq2000 to the reference genome using BWA. I’ve called SNPs using SAMTools and now need to call indels. Under the NGS: Indel Analysis toolset, I see two options: “Filter Indels for SAM” and “Extract indels from SAM.” Since the descriptions are a little vague, I would presume that I start with the “Filter Indels for SAM” first on the BWA SAM file and then run “Extract indels from SAM” on the “Filter” output SAM file (i.e. BWA SAM -> Filter Indels for SAM -> Extract indels from SAM). Is this the correct order? Or would I skip the “Filter Indels for SAM” step (since the BWA SAM file technically already contains indels so there would be no need to filter) and just go straight to “Extract indels from SAM” (i.e. BWA SAM -> Extract indels from SAM)?
I’ve tried both ways and get different results. For example:
1.BWA SAM -> Filter Indels for SAM -> Extract indels from SAM -> gives me 26,417 regions of interest
2.BWA SAM -> Extract indels from SAM -> gives me 93,974 regions of interest
Which way is correct? Any help/info would be greatly appreciated!
Thanks,
David
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David K Crossman -
Jennifer Jackson