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Old 01-14-2013, 10:25 AM   #1
Location: NYC

Join Date: Jun 2010
Posts: 18
Default Plasmid library sequencing-which platform (if any)?

Hi everyone

I was hoping to pick your brains for a second. We have a plasmid library in e coli. We would like to obtain sequences form hundred of individual plasmids, starting from the backbone (meaning from an arbitrary primer) into the inserts.

The classical way to do this is miniprep and sanger sequencing of individual colonies. At the numbers that we would like to do, the price goes well into NGS territory.

I've been trying to find an NGS platform that would provide similar information from a plasmid prep of pooled colonies. But all the methods I've seen involve library prep and introducing an adapter that will serve as the primer for sequencing. Simple shotgun sequencing (by illumina for instance) of such a sample would give us the information, but only <5% of the reads would be informative (the rest would be backbone).

I just know that I haven't looked at all the miriad methods that have been developed. Is there a technology that will give you sequence from an arbitrary primer? Around 400bp would be great, but we could live with less.

Thanks in advance!
zaratieg is offline   Reply With Quote