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Bowtie2 and BWA-MEM not working

Hi, so I have been using Galaxy for genome alignment on two sequences (one forward and other reverse) obtained from Illumina sequencing. The sequences are DNA from exome sequencing. I have realized that when I use the tool "filter quality by data" on my two sequences and then try to align them using either Bowtie2 or BWA-MEM, I get an error message. The error message I get from Bowtie 2 is:

Fatal error: Exit code 1 ()
Tool generated the following standard error:

[bam_sort_core] merging from 16 files and 8 in-memory blocks...

The error I get from BWA-MEM is:

[M::mem_pestat] skip orientation FF as there are not enough pairs
[M::mem_pestat] skip orientation FR as there are not enough pairs
[M::mem_pestat] skip orientation RF as there are not enough pairs
[M::mem_pestat] skip orientation RR as there are not enough pairs
[mem_sam_pe] paired reads have different names: "K00171:532:HLKHHBBXX:5:1101:29254:1754", "K00171:532:HLKHHBBXX:5:1101:29721:1754"

[mem_sam_pe] [mem_sam_pe] paired reads have different names: "K00171:532:HLKHHBBXX:5:1101:28848:1754", "K00171:532:HLKHHBBXX:5:1101:29315:1754"

[mem_sam_pe] paired reads have different names: "K00171:532:HLKHHBBXX:5:1101:28645:1754", "K00171:532:HLKHHBBXX:5:1101:29011:1754"

Why is it the case? Why is it that when I apply the filter tool on sequences, the alignment tools just give up, whereas when I don't filter sequences, they align perfectly. How do I overcome this?

Kind regards, Matt

bowtie2

there is a problem with your fastq files and or the order of the reads in your fastq files . see paired reads have different names (bwa-mem) ; bwa error: paired reads have different names ; etc

for example; you should have the same read names left/right for the following command:

paste  <(gunzip -c in.R1.fq.gz | paste - - - - | cut -f 1)  <(gunzip -c in.R2.fq.gz | paste - - - - | cut -f 1) | grep -F "K00171:532:HLKHHBBXX:5:1101:29254:1754" -m2 -C 3

I use the tool "filter quality by data" on my two sequences

Which tool is that in galaxy? Are you processing your read pairs separately?

No both at the same time and the tool is called "filter by quality"

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