Question: bbmap split paired-end reads back into separated fastq files?
0
gravatar for c.e.chong
4 weeks ago by
c.e.chong0
c.e.chong0 wrote:

Hi,

I am trying to remove human reads from my metagenomic sequences (skin samples).

I have used the command recommended in the bbmap documentation:

bbmap.sh minid=0.95 maxindel=3 bwr=0.16 bw=12 quickmatch fast minhits=2 path=/path/to/hg19masked/ qtrim=rl trimq=10 untrim -ea -Xmx100g in=reads.fq outu=clean.fq outm=human.fq threads=12

My input is Illumina HiSeq 4000 Paired-end, 2x150 bp reads, so I want to split paired-end reads back into separated fastq files .._r1 .._r2.

Is there a way to do this using BBTools?

Thanks in advance!

ADD COMMENTlink modified 4 weeks ago by genomax57k • written 4 weeks ago by c.e.chong0

As shown your command is only using one read file (in=). So unless that file is interleaved there is no chance that out*= files will be interleaved. @ATPoint shows what an interleaved file should look like. Note: Your headers may look different but Read 1 and 2 from one clusters should be one after the other in the file.

ADD REPLYlink modified 4 weeks ago • written 4 weeks ago by genomax57k

Sorry that was a mistake, I have actually used in=r1.fq in2= r2.fq!

Thanks!

ADD REPLYlink written 4 weeks ago by c.e.chong0
1
gravatar for genomax
4 weeks ago by
genomax57k
United States
genomax57k wrote:

Use reformat.sh from BBMap suite. This assumes your clean.fq is interleaved.

reformat.sh in=clean.fq out1=clean_R1.fq out2=clean_R2.fq
ADD COMMENTlink written 4 weeks ago by genomax57k

Thank you very much for your help!

How would I find out if the clean.fq file is interleaved?

ADD REPLYlink written 4 weeks ago by c.e.chong0

Check if two adjacent read pairs always have the same read name. This is an example of an interleaved file from a HiSeq3000:

@HWI-ST-J00104_BSF_0515:8:1101:10003:10036#DMSO_rep2_S43794/1
GGAAGAAAGACAGTCTGTGGCCCTGCCTGGGGACCTACACTGTCTGCTGTAACAGGCTTTCCCTTCATCTCAAGAG
+
AAFFFJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
@HWI-ST-J00104_BSF_0515:8:1101:10003:10036#DMSO_rep2_S43794/2
GGTTGAAAAGTGGCCTCGGTGTTCCAGACCTACCTGGTTCACTTAGCTTTTTCCTCCTTTCCTTCCTTTTATACTC
+
AAFFFJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJFJJJJJJJJJJJJJJJJJJJFJJJJJJJFJJJJJJJJJ
@HWI-ST-J00104_BSF_0515:8:1101:10003:10141#DMSO_rep2_S43794/1
GTGATTCTCTTCAAGGCCACTCCTAAACAACTGTTGAATCCTTGATCCAGGCACCAAGCCCAGAGGTCCCTATCAG
+
AAAFFJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ<FJJJJJJJJJJJJJFJJJJJJJJJJJ
@HWI-ST-J00104_BSF_0515:8:1101:10003:10141#DMSO_rep2_S43794/2
GTCTAGGAGGAAACCAGGCTGTTAACTCCTCAGCAAAGACAAAGGAGGAGTTGGGCGGGGGCAAGACTAACTCCTA
+
AAFFFJJJJJFJJJJJJJJJJJJJJJJJJJJJJJJJFJJJJJJAJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
ADD REPLYlink modified 4 weeks ago • written 4 weeks ago by ATpoint8.1k
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