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  • zillur
    Senior Member
    • Sep 2014
    • 106

    #1

    Bowtie2 using and facing problem

    Hi all,
    I am using bowtie2. Recently I am facing some problem in mapping. I am downloading yeast data from http://www.ebi.ac.uk/ena/data/view/ERX010187. My data showing good in FASTQC but the alignment is not good and finally I am getting nothing. Can anyone help me with this?

    Best Regards
    Zillur

    Zillur-Rahman:1:28:2015 ZILLURRAHMAN$ ls
    ERR029138_1.fastq ERR029138_1.fastq.gz ERR029138_2.fastq ERR029138_2.fastq.gz
    Zillur-Rahman:1:28:2015 ZILLURRAHMAN$ bowtie2 -x ../genome -1 err029138_1.fastq -2 err029138_2.fastq -S eg.sam
    30909942 reads; of these:
    30909942 (100.00%) were paired; of these:
    30909942 (100.00%) aligned concordantly 0 times
    0 (0.00%) aligned concordantly exactly 1 time
    0 (0.00%) aligned concordantly >1 times
    ----
    30909942 pairs aligned concordantly 0 times; of these:
    0 (0.00%) aligned discordantly 1 time
    ----
    30909942 pairs aligned 0 times concordantly or discordantly; of these:
    61819884 mates make up the pairs; of these:
    43136222 (69.78%) aligned 0 times
    557574 (0.90%) aligned exactly 1 time
    18126088 (29.32%) aligned >1 times
    30.22% overall alignment rate
    Zillur-Rahman:1:28:2015 ZILLURRAHMAN$ samtools view -bS eg.sam > eg.bam
    [W::sam_hdr_parse] duplicated sequence 'I'
    [W::sam_hdr_parse] duplicated sequence 'VI'
    [W::sam_hdr_parse] duplicated sequence 'III'
    [W::sam_hdr_parse] duplicated sequence 'IX'
    [W::sam_hdr_parse] duplicated sequence 'VIII'
    [W::sam_hdr_parse] duplicated sequence 'V'
    [W::sam_hdr_parse] duplicated sequence 'XI'
    [W::sam_hdr_parse] duplicated sequence 'X'
    [W::sam_hdr_parse] duplicated sequence 'XIV'
    [W::sam_hdr_parse] duplicated sequence 'II'
    [W::sam_hdr_parse] duplicated sequence 'XIII'
    [W::sam_hdr_parse] duplicated sequence 'XVI'
    [W::sam_hdr_parse] duplicated sequence 'XII'
    [W::sam_hdr_parse] duplicated sequence 'VII'
    [W::sam_hdr_parse] duplicated sequence 'XV'
    [W::sam_hdr_parse] duplicated sequence 'IV'
    Zillur-Rahman:1:28:2015 ZILLURRAHMAN$ samtools sort eg.bam eg.sorted
    [bam_sort_core] merging from 26 files...
    Zillur-Rahman:1:28:2015 ZILLURRAHMAN$ samtools mpileup -uf ../genome.fa eg.sorted.bam | bcftools view -o - > eg.raw.bcf
    [mpileup] 1 samples in 1 input files
    <mpileup> Set max per-file depth to 8000
    Zillur-Rahman:1:28:2015 ZILLURRAHMAN$ bcftools stats eg.raw.bcf
    # This file was produced by bcftools stats (1.1+htslib-1.1) and can be plotted using plot-vcfstats.
    # The command line was: bcftools stats eg.raw.bcf
    #
    # Definition of sets:
    # ID [2]id [3]tab-separated file names
    ID 0 eg.raw.bcf
    # SN, Summary numbers:
    # SN [2]id [3]key [4]value
    SN 0 number of samples: 1
    SN 0 number of records: 0
    SN 0 number of SNPs: 0
    SN 0 number of MNPs: 0
    SN 0 number of indels: 0
    SN 0 number of others: 0
    SN 0 number of multiallelic sites: 0
    SN 0 number of multiallelic SNP sites: 0
    # TSTV, transitions/transversions:
    # TSTV [2]id [3]ts [4]tv [5]ts/tv [6]ts (1st ALT) [7]tv (1st ALT) [8]ts/tv (1st ALT)
    TSTV 0 0 0 0.00 0 0 0.00
    # Sis, Singleton stats:
    # SiS [2]id [3]allele count [4]number of SNPs [5]number of transitions [6]number of transversions [7]number of indels [8]repeat-consistent [9]repeat-inconsistent [10]not applicable
    SiS 0 1 0 0 0 0 0 0 0
    # AF, Stats by non-reference allele frequency:
    # AF [2]id [3]allele frequency [4]number of SNPs [5]number of transitions [6]number of transversions [7]number of indels [8]repeat-consistent [9]repeat-inconsistent [10]not applicable
    # QUAL, Stats by quality:
    # QUAL [2]id [3]Quality [4]number of SNPs [5]number of transitions (1st ALT) [6]number of transversions (1st ALT) [7]number of indels
    # IDD, InDel distribution:
    # IDD [2]id [3]length (deletions negative) [4]count
    # ST, Substitution types:
    # ST [2]id [3]type [4]count
    ST 0 A>C 0
    ST 0 A>G 0
    ST 0 A>T 0
    ST 0 C>A 0
    ST 0 C>G 0
    ST 0 C>T 0
    ST 0 G>A 0
    ST 0 G>C 0
    ST 0 G>T 0
    ST 0 T>A 0
    ST 0 T>C 0
    ST 0 T>G 0

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