No variants found ( have problem in the handbook)
I am new in bioinformatics. Today I was studying variation calling fllowing "the biostars handbook", when I found a bug out of my knowledge. Here is my code which is almost the same with the book chapter 87 : Variant calling example.
# Get the reference sequence.
efetch -db nuccore -format fasta -id $ACC | seqret -filter -sid $ACC > $REF
# Index reference for the aligner.
bwa index $REF
# Index the reference genome for IGV
samtools faidx $REF
# Get data from an Ebola sequencing run.
fastq-dump -X 100000 --split-files $SRR
# Shortcut to read names.
R1=${SRR}_1.fastq
R2=${SRR}_2.fastq
# Align and generate a BAM file.
bwa mem $REF $R1 $R2 | samtools sort > $BAM
# Index the BAM file.
samtools index $BAM
# Determine the genotype likelihoods for each base.
bcftools mpileup -Ov -f $REF $BAM > genotypes.vcf
# Call the variants with bcftools.
bcftools call --ploidy 1 -vm -Ov genotypes.vcf > variants1.vcf
When I view the resulting VCF file in IGV, I got "No variants found":
I can tell that my igv is good, that is I think it's the problem caused by other reasons. Good wishes for you!
• 1,176 views
•
link
0 answers
No answers yet.
Log in to answer this question.
Appears as if you are using the wrong reference to view the data in IGV. Can you confirm that Handbook asks you to use AF08633 as the reference?
I can guarantee it!