CG content and Illumina Sequencing

[David [R]]

Hi,

We are having some problems sequencing DNA regions with a GC% higher than 75 in a bacteria with a GC average content of 67%. We are not completely sure whether the problem comes from the libraries or from the sequencing itself. The former is done with v2 from TruSeq and the latter is done with the v3 from Illumina, with no protocol modifications in either case.

Thanks in advance for your help,

David

[colindaven]

Yep, GC can be nasty. We have seen similar issues.

This is a great paper on the subject which will hopefully solve this issue:

http://www.ncbi.nlm.nih.gov/pubmed/21338519

[David [R]]

Thanks a lot!

Thanks a lot!