This is an old revision of the document!
John St. John's lecture on EULER-SR and Celera; Michael Cusack's lecture on MIRA
campusrocks is broken!
Pog has 2 repeats: ~1k & 1.1k
use makefiles, not shell scripts!
SOLiD data formats:
.csfasta = colorspace with numbers
.de = changes #s to letters (0123 → ACGT) but it’s colors not numbers! very confusing.
.fa is the real basespace
Kevin I mapped newbler to join the contigs found a bug in the python script to map the solid reads. Detected because there were no joining reads for the two that joined the extrachromosomal reads. There was a sign error in one of my tests. Re-did colorspace mapping on newbler5 assembly. May still have a bug since one gap is covered by 10 thousand reads whereas the other side has one that is only covered by 200 reads. Will be looking to see if there is another bug. If you have mate-pair data, it's good to have software to check for correct answers. Pog matepair data, compare to other assembly tools.
SR == Short Reads
Euler-SR is a short-read De Bruijn Graph assembler that can use long reads and mate-pairs.
ran well first time (it ran, at least)
have to run it where you installed it
~2k contigs which create a 2x long genome… suspicious
are contigs overlapping?
check blat_strict_match (blat alignment to reference genome)
look for “Q name” (contigs) which match to the same “T start” positions on the reference genome
answer:yes, appear to overlap a lot – double coverage because they totally overlap
Things to try to improve the run:
- longer k-mers
- increase frequency threshold (help make up for read errors, maybe?)
“Error Correction via threading”
- took reads that “they couldn’t make error free”
- made contigs out of these
- tried to map them back to the “error-free” contigs
- perhaps this is where it went wrong?
Tried to run on just the SOLiD data… started on Sunday, but still running (Wed)
needs qual info (need this from Sanger reads, too)
… so can't run unless you have the .qual files
seemed to have a script to convert Illumina → their format… but not released yet
with 454 data alone: 386 contigs
(newbler: ~40 contigs)
took about 50min
needs datafile named pog_in.[format].fa
sff_extract script to create .qual files
created 30 contigs >=500 (largest contig 640k)
but… upon mapping to the reference genome,
it turns out that while it is making big contigs, it's producing a chimeric assembly, in which the contigs join genomic regions that are not truly adjacent. it’s getting bigger contigs because it’s joining them incorrectly!
this is very bad; worse even than a lot of small contigs