The pcr.seqs will trim inputted sequences based on a variety of user-defined options.
The fasta parameter is required.
mothur > pcr.seqs(fasta=silva.bacteria.fasta)
The oligos file allows you to provide primer information.
mothur > pcr.seqs(fasta=silva.bacteria.fasta, oligos=pcrTest.oligos)
The pdiffs parameter is used to allow differences in the forward primers. pdiffs is maximum number of differences to the forward primer sequence, default=0.
The rdiffs parameter is used to allow differences in the reverse primers. rdiffs is maximum number of differences to the reverse primer sequence, default=0.
The checkorient parameter will look for the reoriented and reverse compliment of the primers if the primers are not found. The default is true.
name && group && count && taxonomy
If you apply pcr.seqs as we have in the previous sections and then attempt to run any downstream commands that need a name, group and taxonomy file, the files will be incompatible since those files will still contain information for sequences that you culled. To get around this you can use the group and name or count option:
mothur > pcr.seqs(fasta=silva.bacteria.fasta, taxonomy=silva.bacteria.rdp.tax)
The ecoli parameter is used to provide a fasta file containing a single reference sequence (e.g. for e. coli) this must be aligned. mothur will trim to the start and end positions of the reference sequence.
start && end
The start parameter allows you to provide a starting position to trim to. The end parameter allows you to provide a ending position to trim from.
mothur > pcr.seqs(fasta=silva.bacteria.fasta, start=1044, end=6500)
The nomatch parameter allows you to decide what to do with sequences where the primer is not found. Default=reject, meaning remove from fasta file. if nomatch=keep, then do nothing to sequence.
The keepprimer parameter allows you to keep the primer, default=false. Note that even if keepprimer=TRUE, mothur will not remove primers if it finds multiple potential matches to a given primer sequence.
The keepdots parameter allows you to keep the leading and trailing .’s, default=true.
The processors parameter allows you to use multiple processors. Default processors=Autodetect number of available processors and use all available.
- 1.25.0 - First Introduced
- 1.28.0 - Added count option
- 1.29.2 - Bug Fix: If file was aligned and both forward and reverse primers were given, reverse primer was not trimmed properly.
- 1.30.0 - Added pdiffs parameter
- 1.30.2 - Bug Fix: For forward primer trimming with aligned sequences and keepdots=t. If the character before the first primer base was a base and not a gap the base was not trimmed. https://forum.mothur.org/viewtopic.php?f=4&t=2209
- 1.31.0 - Added primer to oligos types pcr.seqs can read.
- 1.31.0 - Bug Fix: removing primers from aligned sequences with keepdots=f could result in an unaligned dataset if primers were not found at the same locations. Added gaps to preserve alignment.
- 1.33.0 - Bug Fix: keepdots=f could cause an aligned template to become unaligned. https://forum.mothur.org/viewtopic.php?f=3&t=2653&p=7354#p7354
- 1.38.0 - Adds rdiffs to allow for setting different diffs for the forward and reverse primers.
- 1.40.0 - Rewrite of threaded code. Default processors=Autodetect number of available processors and use all available.
- 1.40.0 - Bug Fix: Trimming extra base when using the start and end parameters. #348
- 1.40.0 - Bug Fix: “name mismatch” error when using paired primers and keepdots=f.
- 1.40.4 - Bug Fix: pcr.seqs and screen.seqs (with no bad reads detected), causing accnos file issue.
- 1.41.0 - Bug Fix: Fixes pcr.seqs file mismatch. #522
- 1.42.0 - Bug Fix: Fixes file extensions of output files in pcr.seqs.
- 1.42.0 - Bug Fix: Fixes pcr.seqs end issue with keepdots=f.
- 1.46.0 - Adds checkorient parameter. Change Includes functions to reorient and reverse unpaired primers. #768
- 1.47.0 - Fixes bug with pcr.seqs If keepdots=f, and start and end are used, then first base was removed by mistake.