I am planning to do Illumina HiSeq sequncing on a relatively small number of exons for 96 individuals (snakes). I am thinking about using the Illumina's Nextera 96-indexing DNA library prep kit for barcoding and prepping the sequencing libraries, in combination with the MySelect kit for custom sequence capture. These 96 barcoded sequence capture libraries will then be pooled for a single lane on the HiSeq2000. Has anyone tried the Nextera kit and the MySelect kit in combination? If so, was it successful? I am most worried about the small starting amount of DNA for the Nextera kit causing targeted exons be missed. Thanks for any advise you can provide.
Location: Ann Arbor, MI, USA
Join Date: Mar 2010
Posts: 5
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Please note that MYselect has been renamed MYbaits. Same exact product, just a new name that better reflect the kit content.
http://www.mycroarray.com/mybaits/MY...hment+kit.html
Hi! I´m planning to use Illumina Nextera 96 indexing library too, and if you already did use it I want to know some data about your experience, what is the size of your exons? and what is the quality of your run, depth and coverage?
Hi Noe, I decided not to use the Nextera kit because I was concerned about the 50 ng starting amount being to small to get good coverage for exon capture. I used Bioo Scientific's Nextflex's DNA sequencing kit with 96 barcodes. Still working on analyzing the data so I don't have numbers for you, but in general coverage and quality looks pretty good. Exon size varied from 60 bp to over 3800 bp. The probes were tiled 60 bp MYbaits designed by MYSelect.
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