what would be the best coverage for "total length (Mb): 4.21561" of a bacterial genome sequenceing?
2
0
Entering edit mode
8.0 years ago
mhmtgenc85 ▴ 50

I have a bacteria on lab and have its reference genome so what would be the best coverage for "total length (Mb): 4.21561" of a bacterial genome sequenceing? and hiseq or miseq?

genome sequencing ngs bacteria hiseq • 2.2k views
ADD COMMENT
0
Entering edit mode
8.0 years ago

That depends on desired or possible read length, downstream application (why sequence it) and repetitive content of the genome.

ADD COMMENT
0
Entering edit mode

I would like to do whole genome sequencing (4,000,000bp) of a bacteria which has a reference sequence. so I won't do de novo assembly I will do reference based assembly. So what is your opinion?

ADD REPLY
0
Entering edit mode

MiSeq should be sufficient to cover a genome like that, MiSeq V3 has ~45M 300bp paired-end reads (depending on cluster density). Just rough calculation: 40 million reads of 300bp => 3000 fold coverage which is an enormous/outrageous over sequencing. You definitely don't need HiSeq for this, but offcourse it depends on what machines you have available and/or the prices of your sequencing provider.

Since your genome is haploid about 5-15 fold coverage should be fine for calling SNPs...

ADD REPLY
0
Entering edit mode
8.0 years ago
GenoMax 141k

Try Illumina's sequencing coverage calculator.

ADD COMMENT
0
Entering edit mode

Tried it. But they actually ask the coverage to me so I don't really understand why :)

ADD REPLY
0
Entering edit mode

Since you have a reference available coverage you want would depend on what you are trying to do as @decosterwouter said above.
If you are looking to call SNP's (or do rearrangement analysis) then 5-10x coverage may be enough. If your strain turns out to differ significantly from the reference (and many strains might depending on how long they have been cultured/in use in your lab) you may need to assemble a new reference for your strain.

ADD REPLY

Login before adding your answer.

Traffic: 2527 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