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  • hugorody
    Junior Member
    • May 2013
    • 9

    #1

    Fasta consensus, SNP call and FreeBayes

    Hi everyone
    I have one question about FreeBayes.

    I'm trying to call SNP from haploid genome with the follow lines:

    ------------------------------------------------------------------------------------------------------------------------
    samtools faidx ref.fasta (index fasta reference)
    samtools view -bt ref.fasta.fai aln.sam > aln.bam (convert bowtie2 alignment *.sam file to bam)
    samtools sort aln.bam aln.sorted (apply sort)
    samtools calmd -Abr aln.sorted.bam ref.fasta > aln.baq.bam
    samtools index aln.baq.bam (index bam files)

    freebayes --min-base-quality 20 -p 1 --fasta-reference ref.fasta aln.baq.bam > aln.vcf (create the VCF file)

    bcftools view -D -1 -Sb aln.vcf > aln.bcf (convert VCF to BCF)

    samtools mpileup -uf ref.fasta aln.baq.bam | bcftools view -vcg -S aln.vcf - | perl vcfutils.pl vcf2fq > aln.consensus.fq (create a consensus sequence with SNPs)
    ------------------------------------------------------------------------------------------------------------------------

    The problem is:
    The file aln.consensus.fq do not show all the bases that match to reference sequence. Instead it, the fq file just show the SNPs between "n" like this: nnnnnnncnnnnnannnnttnnnnnnnnnannnnn.

    Someone can help me to solve this?
    Thanks

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