Hello everybody,
I'm interesting for the mitochondrial assembly from several samples of sheep reads.
Based on the post "Tutorial: Reference Assembly - Mapping Reads To A Reference Genome" I have mapped my reads with the sheep reference by using bowtie2 and stampy.
I have generated the bam files, I extracted the reads that are mapped onto the mitochondrial by using samtools and after that again with samtools and the following command I generated the consensus (fq)
samtools mpileup \
-E \
-uf oviAri3.fa \
B2_ACAGTG_L001_PE_stampy.sorted.chrM.bam | \
bcftools view -cg - | \
vcfutils.pl vcf2fq > B2_ACAGTG_L001_PE_stampy_chrM_cons.fq
or
samtools mpileup \
-E \
-uf oviAri3.fa \
B2_ACAGTG_L001_PE_bwt.sorted.chrM.bam | \
bcftools view -cg - | \
vcfutils.pl vcf2fq > B2_ACAGTG_L001_PE_bwt_chrM_cons.fq
for bowtie output.
After that, by using the seqret from emboss package I converted the fq to fa.
Comparing the two fa files with the reference, Im seeing that there are some differences. Bowtie assembly has some regions with n's, stampy has only 10 n's at the beginning and a little bit more differences in some bases. Of course the most of the differences with the reference are the same.
How can I identify which assembly is better?
Is there any other step that I have to follow and I didn't do it?
Since I am really new in that field, I will appreciate for any help.
Thank you very much in advance,
Vasilis.
assembly
mitochondria
samtools
consensus