Alignment Error in BWA: The sequence "lcl|" was not found
0
0
Entering edit mode
3.1 years ago
gt ▴ 30

I have aligned DNA-seq data to the following reference genome: https://www.ncbi.nlm.nih.gov/nuccore/U00096 I keep getting the following output in my error file when I try to run the bcftools mpileup command for variant calling:

[E::faidx_adjust_position] The sequence "lcl|" was not found
[E::faidx_adjust_position] The sequence "lcl|" was not found
[E::faidx_adjust_position] The sequence "lcl|" was not found
[E::faidx_adjust_position] The sequence "lcl|" was not found
[E::faidx_adjust_position] The sequence "lcl|" was not found
[E::faidx_adjust_position] The sequence "lcl|" was not found
[E::faidx_adjust_position] The sequence "lcl|" was not found
[E::faidx_adjust_position] The sequence "lcl|" was not found
[E::faidx_adjust_position] The sequence "lcl|" was not found

I have these same error lines for tons of rows. I know in my .fa file which I downloaded I have the following first line

>NC_000913.3 Escherichia coli str. K-12 substr. MG1655, complete genome

Do I have a missing string here or something at the beginning? I did not touch the file when I downloaded it so I just assumed the alignment would work. This is the command I am using to align:

bwa mem -t 180 ecoli R1_001.fastq R2_001.fastq > sample.sam

This is the command I am using to find the variants:

bcftools mpileup -d 500 -Ou -f GCF_000005845.2_ASM584v2_genomic.fa sample.bam | bcftools call -mv -Ob -o sample_calls.bcf

which I took directly from the Github page.

"ecoli" is the index I built using the .fa and .gtf files from the same website. Any help appreciated here! Important note: I have not included my file paths here for privacy reasons.

bwa DNA-Seq alignment • 735 views
ADD COMMENT
0
Entering edit mode

Just to be sure you downloaded fasta format of the genome sequence correct? Do you just see only one line when you do grep "^>" your_fasta_file or additional lines?

ADD REPLY

Login before adding your answer.

Traffic: 2937 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6