It's no available, the web page cannot be open. http://genomics.jhu.edu/software/FLASH/index.shtml
Hi,
How can I merge two paired end fastq (R and L) to give a single fastq file ? For information, the sequencing run is 72 bp long and it contains a majority of small RNA (miRNA,...) so a lot of paired end reads will overlap.
For example here's two paired reads :
@HWUSI-EAS529:41:FC62YHFAAXX:8:1:7969:1330 1:N:0:GCCAAT
CTACGAAAGGGCACTTGGAATTCTCGGGTGCCAAGGAACTCCAGTCACGCCAATATCTCGTATGCCGTCTTCTGCT
+
IIIIIIIHIIHIIIIIIHHIIIHGIIIIEIIIIIIEIIHIIIIIIIIIIIHIIIIIBHIHIIHGIGIEGHHEGEEH
@HWUSI-EAS529:41:FC62YHFAAXX:8:1:7969:1330 2:N:0:GCCAAT
AGTGCCCTTTCGTAGGATCGTCGGACTGTAGAACTCTGAACGTGTAGATCTCGGTGGTCGCCGTATCATTAAAAAA
+
IIIIIIIIIIIIIIIIIIIIIIIDHIGIIIHIIIGHGIIIIIIIHHIHIIIIIIIIIHIIIIIIIIHIIGIIIIHI
I find the adapter in the first one:
Code:
EMBOSS_001 1 CTACGAAAGGGCACTTGGAATTCTCGGGTGCCAAGGAACTCCAGTCACGC 50
|||||||||||||||||||||
EMBOSS_001 1 ---------------TGGAATTCTCGGGTGCCAAGG-------------- 21
EMBOSS_001 51 CAATATCTCGTATGCCGTCTTCTGCT 76
EMBOSS_001 22 -------------------------- 21
but not in the second one
But I effectively found the overlap between the right read and the left read (using the reverse complement of it)
EMBOSS_001 1 -------------------------------------------------- 0
EMBOSS_001 1 TTTTTTAATGATACGGCGACCACCGAGATCTACACGTTCAGAGTTCTACA 50
EMBOSS_001 1 -----------CTACGAAAGGGCACTTGGAATTCTCGGGTGCCAAGGAAC 39
|||||||||||||||
EMBOSS_001 51 GTCCGACGATCCTACGAAAGGGCACT------------------------ 76
EMBOSS_001 40 TCCAGTCACGCCAATATCTCGTATGCCGTCTTCTGCT 76
EMBOSS_001 77 ------------------------------------- 76
So my question is, how can I merge the two fastq files to produce a single fastq file?
Thanks,
N.
8 answers
Maybe this program is suited for you: http://www.cbcb.umd.edu/software/flash/
It's here now: http://ccb.jhu.edu/software/FLASH/
There is a decent program called stitch: https://github.com/audy/stitch
I wrote a script called mergePairs that is very sensitive and incredibly slow: http://code.google.com/p/standardized-velvet-assembly-report/source/browse/trunk/mergePairs.py
Either use Galaxy or use the single scripts
http://hg.notalon.org/galaxy/galaxy-central/src/7d9bb95caaa7/tools/fastq
HTH!
uhhh which script?
If you wish to trim adapters and merge in a single step, you can the leeHom, we use it mainly to reconstruct ancient DNA sequences but it has broader uses as well:
http://nar.oxfordjournals.org/content/42/18/e141
Click here for the Website of the repository
It use a Bayesian maximum a posteriori approach that considers quality scores for both the adapter determination and the merging part.
There is BBMerge, which is ""designed to merge two overlapping paired reads into a single read. For example, a 2x150bp read pair with an insert size of 270bp would result in a single 270bp read"": http://jgi.doe.gov/data-and-tools/bbtools/bb-tools-user-guide/bbmerge-guide/
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Hi, I see that you had a similar case like me, so probably you can help me :)
As I always do the miRNA analysis in single end I'm confused how to proceed when I have paired-end? Can you recommend me how to clean the reads and have them ready for analysis, particularly I cannot understand how and what is the relation of the reverse-compliment miRNA sequence in R2 read to the R1 set?
In summary my R1 read is containing 100nt - miRNA+barcode+smallRNAadapter+another adapter+polyA
my R2 is containing miRNA (reversed compliment to R1) + long adapter (or linker) + polyA
Thanks for any help in advance!
Hi,
So for an exact answer to this problem.
The
R1.fqare the forward reads and theR2.fqare written in reverse-complement.For example, if I want to create a single file from reads in R1.fq and R2.fq, I have to do "reverse-complement" of reads in R2.fq?
Am I right?
Thank you
no response for this problem?