wiki:SOPs/mapping

Version 4 (modified by gbell, 12 years ago) ( diff )

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Mapping short reads

Regular mappers

These mapping tools are useful for reads of DNA origin that should map to a continuous stretch of genomic DNA. Some of these tools can tolerate short indels but they're not designed for reads that span a splice junction

One may choose between bowtie version 1 (faster but ignores indels) and bowtie version 2 (slower but allows indels). For a feature comparision, see How is Bowtie 2 different from Bowtie 1?

bowtie version 1

Sample command:

bsub "bowtie  -k 1 -n 2 -l 70 --best --sam --solexa1.3-quals /nfs/genomes/mouse_gp_jul_07_no_random/bowtie/mm9 s_7.txt ../s7_mm9.k1.n2.l36.best.sam"

The parameters used on the sample command are:

  • -l/--seedlen <int> seed length for -n (default: 28) -- Set to longest possible length of high-quality bases. Use the FastQC output to determine length of high-quality positions.
  • -n/--seedmms <int> max mismatches in seed (can be 0-3, default: -n 2)
  • -k <int> report up to <int> good alignments per read (default: 1) -- If you want only uniquely mapped reads, however, also use '-m 1' to ignore multi-mapped reads
  • --solexa1.3-quals (if input quals are from GA Pipeline ver. >= 1.3) See the table at the top of FastQC output to identify the "encoding" scale
  • --best (in the case of multi-mapped reads, keep only the best hit(s))
  • --sam to get SAM output format (which is the best format for downstream analysis)

To see other parameters log into tak and type bowtie

bowtie version 2

Splice-aware mappers

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