I think you can do most of what you want (following the bowtie alignment step) with samtools, using samtools sort, index, and then mpileup.
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Getting transcriptome sequence from RNA-seq reads in FASTA
Hello,
So, from Ensembl FTP, we can download a transcriptome file which is a FASTA file containing the header info, which is transcript name, chromosome position, etc. and the dna sequence itself. On the other hand, I have a FASTQ file from RNA-seq experiment.
What I want to do is, generate the FASTA file like Ensembl transcriptome. I think I read this is called consensus FASTA.
What I imagine the step to generate this is like this:
1. Align the reads to the transcriptome reference, we get SAM/BAM
2. Assemble the SAM/BAM according to coordinate
3. Solve the occurence of SNP and indel
4. Generate FASTA file with header information and sequence assembled from step 3
For step 1, I know I can use bowtie2. For step 2, I don't know the tools but I think I can write my own program. The problem is step 3. I don't know how.
In that case, probably you can suggest me well known pipeline to do this because I think this is a general things to do.
What do you suggest for that? Thank you for your reply.Last edited by barbarian; 03-03-2016, 08:32 PM.
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CRISPR/Cas9 sparked the gene editing revolution for both research and therapeutics.1 But this system still showed severe issues that limited its applications. The most prominent were the heavy reliance on PAM sequences, delivery limitations, double-stranded breaks that prompt unintended edits and cell death, and editing inefficiency (both in targeting and in knock-in reliability).
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