[Bioperl-l] Primer maps?
Harry Mangalam
hjm at tacgi.com
Thu Feb 16 17:02:07 UTC 2006
A bit off the bioperl topic - if you must have bioperl, ignore this (or just
system() wrap the command) - but you can do exactly this mapping and in-line
translation with a thing I wrote called tacg - you make a GCG-formatted file
of primers ie for each pattern you need a line like:
; Top Bottom
;Name Offset Recognition Pattern Offset ! comments
primer1 0 tcgggywmkkgg 0 ! ...
primer2 0 gcttggctgaggag 0 !
.
.
.
Obviously the offsets can be set to 0 for non REs.
There's no limit to the number of primer patterns (tho I think there's a
compiled-in limit of 30 chars in the pattern - easily changed in header), no
limit to amount of seq searched, handles degeneracies, searches at ~4Mbases/s
on a 2G opteron (120 patterns).
Also does searching with errors (slowly) and regex's (at pcre speeds), and
matrices. Other neat stuff, too.
The output is sort of as you describe - replace the RE names with your primer
labels and you'll have it.
6 frame xl with 3 letter abbrievs.
BsrGI BsrGI AflII DraI
\ \ \ \
121 gtgtgtatttgtacactttgtacacttaagacctacacatttcattgtgtttaaattatt 180
3453 cacacataaacatgtgaaacatgtgaattctggatgtgtaaagtaacacaaatttaataa 3512
^ * ^ * ^ * ^ * ^ * ^ *
1 ValCysIleCysThrLeuCysThrLeuLysThrTyrThrPheHisCysValTerIleIle
2 CysValPheValHisPheValHisLeuArgProThrHisPheIleValPheLysLeuLeu
3 ValTyrLeuTyrThrLeuTyrThrTerAspLeuHisIleSerLeuCysLeuAsnTyrTyr
4 HisIleGlnValSerGlnValSerLeuTerValValAsnTerGlnThrTerIleIleVal
5 ThrTyrLysTyrValLysTyrValTerArgSerCysMetGluAsnHisLysPheTerTer
6 HisThrAsnThrCysLysThrCysLysGlyLeuValCysLysMetThrAsnLeuAsnAsn
or 3 frames with 1 letter abbrievs
BsrGI BsrGI AflII DraI
\ \ \ \
121 gtgtgtatttgtacactttgtacacttaagacctacacatttcattgtgtttaaattatt 180
3453 cacacataaacatgtgaaacatgtgaattctggatgtgtaaagtaacacaaatttaataa 3512
^ * ^ * ^ * ^ * ^ * ^ *
1 V C I C T L C T L K T Y T F H C V * I I
2 C V F V H F V H L R P T H F I V F K L L
3 V Y L Y T L Y T * D L H I S L C L N Y Y
read more at tacg.sf.net or reply to me for the latest docs and version - have
to admit the sf site is a bit moldy.
hjm
On Wednesday 15 February 2006 13:20, Michael Coyne wrote:
> Hello all --
>
> I'm having a devil of a time figuring out how to make restriction maps
> using BioPerl. What I'm going for is output similar to GCG's map program,
> but instead of using a set of defined restriction enzymes, I'd like to use
> a set of primers, to create a primer map rather than a restriction map. I
> do not need a table of restriction enzymes that cut or don't cut (or
> primers that match or don't match, in this case), but an honest-to-goodness
> map, something like:
>
> FKP-5->
> |
> CGTTCTATCGATATGGGTGCTATGGAAATAGTATCTACGTTTGATGAATTGCAAGATTAT
> 1921 ---------+---------+---------+---------+---------+---------+ 1980
> GCAAGATAGCTATACCCACGATACCTTTATCATAGATGCAAACTACTTAACGTTCTAATA
>
> a M E I V S T F D E L Q D Y -
>
> I also need translations of orfs, but I can use GenBank files as input to
> the program and thus the CDS translations are already there, so I'm
> guessing that shouldn't be too hard.... How does one create such a map
> using the BioPerl modules?
>
> There are intriguing indications out there that such a thing is possible
> (e.g. the Bio::Map:: * and Bio::Restriction:: * modules), but I can't find
> a single example of code that creates such a basic, bread-and-butter thing
> as a restriction map with orf translations. The documentation to these
> modules is fairly useless to me, consisting mostly of internal methods and
> function prototypes. Perhaps my skills as a Perl programmer are to blame,
> but a clear example of how a map like this is constructed would be a big
> help.
>
> Right now, I'm generating primer maps with system calls to EMBOSS's remap,
> pointing it at a file of primer sequences rather than a file of restriction
> enzyme sequences, but the results are less than desired. I'm considering
> trying to adapt tacg 4.1.0 or sequence extractor 1.1 web-based code to my
> needs, but this seems like a lot of work for an operation I suspect is
> possible in BioPerl.
>
> Any help greatly appreciated...
>
> Mike
>
> ---------------------------------------------------------------------
> //=\ Michael J. Coyne phone: (617) 525-7820
> \=// Channing Laboratory FAX: (617) 264-5193
> //=\ EBRC, Room 617
> \=// 221 Longwood Avenue email:mcoyne at channing.harvard.edu
> //=\ Boston, MA 02115 mjcoyne at comcast.net
> \=//
> ---------------------------------------------------------------------
--
Cheers, Harry
Harry J Mangalam - 949 856 2847 (vox; email for fax) - hjm at tacgi.com
<<plain text preferred>>
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