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README
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This repository contains code for finding discriminatory primers among genomes
of interest. Please refer to the following:
CITATION (method and application to human bacterial pathogens, sub-serotype resolution):
Pritchard L, Holden NJ, Bielaszewska M, Karch H, Toth IK (2012) "Alignment-Free
Design of Highly Discriminatory Diagnostic Primer Sets for Escherichia coli
O104:H4 Outbreak Strains." PLoS ONE 7(4): e34498. doi:10.1371/journal.pone.0034498
CITATION LINK:
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0034498
or
CITATION (application to plant pathogens, species-level resolution)
Pritchard L, Humphris S, Saddler GS, Parkinson NM, Bertrand V, Elphinstone JG,
Toth IK (2013) "Detection of phytopathogens of the genus Dickeya using a PCR primer
prediction pipeline for draft bacterial genome sequences." Plant Pathology, 62, 587-596
doi:10.1111/j.1365-3059.2012.02678.x
CITATION LINK
http://onlinelibrary.wiley.com/doi/10.1111/j.1365-3059.2012.02678.x/full
DEPENDENCIES:
Any of the following Dependencies can be used for running the
'find_differential_primers.py' pipeline:
>> biopython : v1.57 - v1.61
Git Hub version : https://github.com/biopython/biopython
Older Releases (scroll to bottom): http://biopython.org/wiki/Download
>> bx-python : current Hg (Mercurial) version works as of June 19, 2013
Hg version : https://bitbucket.org/james_taylor/bx-python/src
>> EMBOSS (for eprimer3) : v6.30, v6.31, v6.4.0
http://emboss.sourceforge.net/download/
>> primer3 : v1.1.4
http://primer3.sourceforge.net/releases.php
>> prodigal : v1.20
http://code.google.com/p/prodigal/downloads/list?can=1&q=&colspec=Filename+Summary+Uploaded+ReleaseDate+Size+DownloadCount
>> BLAST / BLAST+ : v2.2.22 / v2.2.22+ (Other versions may work)
http://blast.ncbi.nlm.nih.gov/Blast.cgi?CMD=Web&PAGE_TYPE=BlastDocs&DOC_TYPE=Download
BASIC USE:
If you do not have complete genome sequences use the 'stitch_six_frame_stops.py'
script to stitch together the contigs of a single genome (contained within one
multi-FASTA formatted file). The script does this by stitching the contigs with
the following sequence 'NNNNNCATTCCATTCATTAATTAATTAATGAATGAATGNNNNN'. This
sequence contains start and stop codons in all frames. Remember the default
workflow is to only find primers within CDS regions.
Once all genomes have been stitched together, make a config file similar
to the example within the 'O104_primers_5.conf' file.
Finally, run 'find_differential_primers.py'.
FURTHER INFORMATION:
Read the comments contained within the top of each '*.py' file as well as the
Supporting Information ('Methods S1' document) of the above CITATION.
CONTRIBUTORS
============
Leighton Pritchard https://github.com/widdowquinn
Benjamin Leopold https://github.com/cometsong
Michael Robeson https://github.com/mikerobeson