Prediction pipeline
From Crop Genomics Lab.
1. Whole reads were mapped against reference genome
2. Both unmapped reads retrieve using following command
samtools view –hb –f 12 –F 256 input.bam > output.bam |
3. Make fastq file in bam file using bam2fastq
4. Denovo assemble of both unmapped reads using AbySS with k-mer = 51 and q-value = 20 : U-contig
5. Denovo assemble of whole sequencing reads using AbySS with k-mer = 51 and q-vale = 20 : W-contig
6. BLASTN between W-contigs and U-contigs longer than 2k with e-value threshold, 1e-100
7. Retrieve W-contigs which contain full length U-contigs on mid-region, not point-region.
8. BLASTN between W-contigs and G. max reference sequence with e-value threshold, 1e-100
9. Indel candidates were selected by followed condition:
- * Indel candidate pairs have homologies with G. max on same chromosomes
- * Indel candidate pairs have homologies which have same direction
- * homologous region of G.max have to smaller than 10k
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