Hello. I need to get information on the genomes belonging to the Pangolin lineages of coronavirus. I have one reference strain as fasta file, and two strains as fastq file. I filtered them, aligned them to the reference and did variant calling. My advisor said that I MUST assemble the genomes (I don’t understand what this means to him). But the goal is to find information about two strains on https://cov-lineages.org/resources/pangolin.html. I guess I could do this by getting fasta files from VCF with accepting variant calling, but don’t know how to do it.
I would greatly appreciate any help.
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Neil •
You can generate consensus sequence using BBTools like this:
bbmap.sh in=reads.fastq ref=reference.fasta out=mapped.sam
consensus.sh in=mapped.sam ref=reference.fasta out=consensus.fasta
I suggest that you learn what it means to "Assemble a genome". You will not satisfy your advisor if you do not do that, so why bother with hypotheticals? It's not like "Assemble a genome" has some special meaning to your advisor; it is something you should understand at least the basics of if you have a degree in anything related to genetics. You work with VCF files, but VCF files are not actually useful without genomes. It is important to learn what VCF files are, what they mean, and how to generate them. Anyone can filter them on a 0.1234 cutoff for column 7, but the purpose of a bioinformatician is to understand what column 7 means and why 0.1234 is a good cutoff.
See this tutorial: Generating consensus sequence from bam file
Following this tutorial will not give an assembly. Instead, it will give an ALT-genome. The function is called "consensus" but this is in fact a misnomer.
But that's not an assembly...
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First, I agree with Pierre you are not trying hard enough to follow up. I thought I had given extensive advice already. If your supervisor wants an assembly they likely mean de novo assembly. Anyway, why don't you just ask them what they want? Just using a tool because it can produce some output in a certain format without understanding what it does is the kind of black-box bioinformatics that leads to more problems than it solves.