Thanks Devon;
I get one more error while running the trim_galore command. Could you please help me with it?
My error:
perl trim_galore --paired --phred333 --q 20 --a AGATCGGAAGAGC -stringency 5 -e 0.1 --t --r1 35 --r2 35 SRR364313_1.fastq SRR364313_2.fastq
No quality encoding type selected. Assuming that the data provided uses Sanger encoded Phred scores (default)
Writing report to '--paired_trimming_report.txt'
SUMMARISING RUN PARAMETERS
==========================
Input filename: --paired
Trimming mode: single-end
Trim Galore version: 0.3.7
Quality Phred score cutoff: 20
Quality encoding type selected: ASCII+33
Adapter sequence: 'AGATCGGAAGAGC'
Maximum trimming error rate: 0.1 (default)
Minimum required adapter overlap (stringency): 1 bp
Minimum required sequence length before a sequence gets removed: 20 bp
Writing final adapter and quality trimmed output to --paired_trimmed.fq
>>> Now performing quality (cutoff 20) and adapter trimming in a single pass for the adapter sequence: 'AGATCGGAAGAGC' from file --paired <<<
open3: exec of cutadapt -f fastq -e 0.1 -q 20 -O 1 -a AGATCGGAAGAGC --paired failed at trim_galore line 507.
How did you install them? Are you facing errors while installing them?
Yes, I have a problem in installing the cutadapt software itself.
hi..
@Devon Ryan...
please tell me that
can I use this file as a reference genome for Tophat since they have mentioned that word as Variant ??
please check the link given bellow:
ftp://ftp.ncbi.nlm.nih.gov/sra/reports/Assembly/GRCh37-HG19_Broad_variant/Homo_sapiens_assembly19.fasta
This should be a new question, but there's no reason you shouldn't be able to use that. Just keep in mind that the annotation file you might get from elsewhere may or may not match it.
thanks Ryan:
but one thing I didn't understand what is the deference between the UCSC fasta file as a reference and this file... as a reference file will be better...and what about the Variant they have mentioned in root directory ???
"Difference", not "deference".
It's unclear exactly what the differences are. At least the b37 reference from Broad has a slightly different sequence and possibly some different contigs in some cases. You'd have to either ask someone from the Broad (they have documentation about this somewhere) or just look yourself by comparing the sequences. In any case, unless you're calling variants on the same samples, then you might as well just use either the UCSC or Ensembl references.