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Old 06-01-2018, 11:09 AM   #61
rajarapupriya
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Thanks for your quick reply. I ran a test run with both the references in the same command.
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Old 08-20-2018, 06:59 AM   #62
kcamnairb
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Hi Brian,

I'm trying to use bbsplit to separate rnaseq reads from two mixed fungal samples. I'm using the individual transcriptomes as references. I was getting some unexpected results. It seemed that more reads were unambiguously mapping to the reference that is listed first, so I swapped the order of the references and the results changed dramatically. I have ambiguous2=toss, but it seems like it's still using the first best site. Below are my commands and refstats output. Is there anything I'm doing wrong?

Thanks,
Brian
Code:
bbsplit.sh ref=53.fasta,17.fasta \
        in=53_30_r1_S7_R1_001.fastq.gz in2=53_30_r1_S7_R2_001.fastq.gz \
        out_17=map17_53_30_r1_S7_R#_001.fastq.gz \
        out_53=map53_53_30_r1_S7_R#_001.fastq.gz \
        refstats=53_30_r1_S7.stats ambiguous2=toss

#name	%unambiguousReads	unambiguousMB	%ambiguousReads	ambiguousMB	unambiguousReads	ambiguousReads
53	41.51013	1625.01508	57.30665	2219.25878	11241396	15519266
17	1.13394	44.03152	57.30665	2219.25878	307084	15519266        
        
bbsplit.sh ref=17.fasta,53.fasta \
        in=53_30_r1_S7_R1_001.fastq.gz in2=53_30_r1_S7_R2_001.fastq.gz \
        out_17=map17_53_30_r1_S7_R#_001.fastq.gz \
        out_53=map53_53_30_r1_S7_R#_001.fastq.gz \
        refstats=53_30_r1_S7.stats2 ambiguous2=toss

#name	%unambiguousReads	unambiguousMB	%ambiguousReads	ambiguousMB	unambiguousReads	ambiguousReads
53	21.37940	838.36051	67.54242	2623.22348	5789774	18291224
17	11.02890	426.72088	67.54242	2623.22348	2986746	18291224

Last edited by GenoMax; 08-20-2018 at 09:03 AM.
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Old 10-08-2018, 02:48 AM   #63
phuongbigbig
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Default Contamination from human genome?

Hi,

I am working on non-model fish RNA-seq data, I am considering remove human contamination from reads, is this feasible since there is number of orthologs between human and fish?
Is there any recommendation regarding choice of "-minratio" for this case? It seems that 0.56 maybe too low? (I don't have reference genome for this non-model fish, by the way)

P.s: I think there should be different usage strategy of sensitivity or specificity for the case of binning (having 2 reference, i.e host vs contaminant, both have comparative alignment score to judge) AND for the case of decontaminating (only have the reference of contaminant, judgement only based on alignment to contaminant reference).

Thank you very much for your suggestion !
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