Circularization of Large Target Fragments

[HeidiJTP]

Hi all,

I would like to target a ~10 kb region for NGS. I was looking at a paper by Myllykangas et al. (http://www.biomedcentral.com/1472-6750/11/122/) which circularizes a target region and prepares it for Illumina sequencing, using very short (~40 bp) fragments.

I have other reasons for wanting to circularize the sequence and keep it intact prior to sequencing, and was wondering if this is possible for such a large fragment. Any suggestions/references for protocols to target and circularize large fragments would be greatly appreciated.

[krobison]

Mate pair library construction involves such circularization; this paper use Cre-Lox for the circularization step might be a good starting point

http://www.ncbi.nlm.nih.gov/pubmed/22253722.1

[HeidiJTP]

Thanks krobison, I appreciate the suggestion and will look into this paper and similar methods for ideas. By the way, I have also found your blog very useful for staying up-to-date on the latest sequencing technology.