I got it, thanks very much.
hi,
I found there is an unrelated row, not the header, existing in my SAM file, which produced by BWA mem. like the error below ([M::process] read 397352 sequences (120000304 bp)...[M::mem_process_seqs] Processed 397352 reads in 815.105 CPU sec, 67.779 real sec):
SRR5439471.393892 0 chr3 25046051 60 147M155S * 0 0 ATCATGTAGTCTGCCCGGCTCTTTTGGTCCTAGATCAGAAAGAGGCTTTTGCAAACCCACCATTTCTGGGGTTGAATGGGAAATTAGGGAAAGAAAATGAGGTCAAGAAAGGCATGGAGAAAAAAACCGCTGGGCCTGTTGGCCTGAACGGTTCTTGAGAACTGCCATCACTTGTAGCTTCAAATCAGCTGCCAAAAATAGTGGAAAGACTAATTGTTATTTCAGGTATAAAGAAACAGCAACGGTGGACCGATGATCATCGGTCCACCGTGAACTGCCTGAGTTACTTGTCCCTGCCTCTG HHHHHHHHGGGGGGGGHHHEHHHEHHHHGHHHHHHGHHHHHHHHHEFEHHHGGFAGEFGGHHFHHGGGGHHHHHHHHHHHHHHHHHHGHHHHHHHHHHHHHHHHHHGHHHHHHHGHHHHGGGGGEEGGHGFGGGGCGGCBBFFFFFBABBBBBBBBFF4AFFFGGGGGGGGGGHHHHHHHGGHHHHHHHBGHGHHGHGHHHHHHBGHHHHHHHFFHHHHHHHHHGGHHHHGHHGHFHHHHHHHEGEGFHHHGGGGGEFHGHEFEEGHHHGHGGGHBFHHHF2FGHHHHHHHHHHHHHHHHHF NM:i:0 MD:Z:147 AS:i:147 XS:i:0 SA:Z:chr1,183484475,-,61S90M151S,60,0;chr2,171679085,+,271S31M,60,0;
SRR5439471.393892 2064 chr1 183484475 60 61H90M151H * 0 0 TGCTGTTTCTTTATACCTGAAATAACAATTAGTCTTTCCACTATTTTTGGCAGCTGATTTGAAGCTACAAGTGATGGCAGTTCTCAAGAA HHHHHFHGHHGHHHHGGHHHHHHHHHFFHHHHHHHGBHHHHHHGHGHHGHGBHHHHHHHGGHHHHHHHGGGGGGGGGGFFFA4FFBBBBB NM:i:0 MD:Z:90 AS:i:90 XS:i:19 SA:Z:chr3,25046051,+,147M155S,60,0;chr2,171679085,+,271S31M,60,0;
SRR5439471.393892 2048 chr2 171679085 60 271H31M * 0 0 GAACTGCCTGAGTTACTTGTCCCTG
[M::process] read 397352 sequences (120000304 bp)...[M::mem_process_seqs] Processed 397352 reads in 815.105 CPU sec, 67.779 real sec
CCTCTG GGGHBFHHHF2FGHHHHHHHHHHHHHHHHHF NM:i:0 MD:Z:31 AS:i:31 XS:i:0 SA:Z:chr3,25046051,+,147M155S,60,0;chr1,183484475,-,61S90M151S,60,0;
SRR5439471.393893 16 chr3 25046051 60 151M151S * 0 0 ATCATGTAGTCTGCCCGGCTCTTTTGGTCCTAGATCAGAAAGAGGCTTTTGCAAACCCACCATTTCTGGGGTTGAATGGGAAATTAGGGAAAGAAAATGAGGTCAAGAAAGGCATGGAGAAAAAAACCGCTGGGCCTGTTGGCCTGAGCACGCCAACCACCCACCTCGGTCTCCCAAAGGGCTGGAACGGTGGACCGATGATCATCGGTCCACCGTTCTGTTACTTAGGGAAATTACAAGCATATTAAGTTCAGCTGCCTTAGCAAAGCTTCAGAACTTCTTAAGGTTTAATATTTTTATTA FEGHHGHHHEGGGGGGHHEGGEHHGGGBHHFHHHHGHHGHHHHHG3HHHHHGGGGGGFHHHHHFGGGGGHHHHHHHHHHHHHHHHGGHHHHHHHGHHHHHHGHHHHHHHHHHGHHHHHHGGGGGGEGGHGGGGGGGGGGCBFF4FFAABBBABCCCCCCCCCCGGGGGGGGGGHHHHHHFGGHGGHHHFGBFFGHHGGGGHHHHHHHGGGGHHHGGEHGHHGHHHHHHHHHHHHHHHHHHHHHHHHHHHHGHHHHHHHHHHHHHHHHHHGHHHHHHHHHHHHGHHHHHHHHHHHHHHHGHHG NM:i:0 MD:Z:151 AS:i:151 XS:i:0 SA:Z:chr3,25045723,-,216S86M,60,0;chr4,169910618,+,115S36M151S,37,0;
SRR5439471.393893 2064 chr3 25045723 60 216H86M * 0 0 TCTGTTACTTAGGGAAATTACAAGCATATTAAGTTCAGCTGCCTTAGCAAAGCTTCAGAACTTCTTAAGGTTTAATATTTTTATTA EHGHHGHHHHHHHHHHHHHHHHHHHHHHHHHHHHGHHHHHHHHHHHHHHHHHHGHHHHHHHHHHHHGHHHHHHHHHHHHHHHGHHG NM:i:0 MD:Z:86 AS:i:86 XS:i:20 SA:Z:chr3,25046051,-,151M151S,60,0;chr4,169910618,+,115S36M151S,37,0;*
And my alignment command is
nohup bwa mem -t 12 GRCh38.bwa VEGFA_site_1.fastq.gz > VEGFA_site_1_output.sam &
Was my command incorrect? Thanks for your help!!!
3 answers
Well, there is your problem. If you use nohup then STDOUT and STDERR get combined in the output file. Either don't use nohup or use the -o option as mentioned by Bastien Hervé
Please use the option -o to output your sam file and not redirecting to standard output using >. In your case you have BWA verbose in your sam file, caused by redirection
bwa mem -t 12 GRCh38.bwa VEGFA_site_1.fastq.gz -o VEGFA_site_1_output.sam
If you must use nohup and there is no -o FILE option call bwa from an inner bash call.
nohup bash -c 'bwa mem -t 12 GRCh38.bwa VEGFA_site_1.fastq.gz > VEGFA_site_1_output.sam' &
That should take care improper redirection of stdout.
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hum... stderr is mixed with stdout with this command (?). was it really your only command ?
Did you align with
nohup?yeah, actually the command is "nohup bwa mem -t 12 GRCh38.bwa VEGFA_site_1.fastq.gz > VEGFA_site_1_output.sam &", but there is nothing in nohup.out file.