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vl80 11-10-2012 05:36 PM

Short PCR product sequencing with Miseq
 
Is it possible to omit the fragmentation step of the library preperation, proceed with the rest of the steps and sequence PCR products of 90 - 450 bp using Miseq?
I am just theorising as I have no prior experience in this but need to plan a way to sequence short PCR products woth Miseq. Any inputs are welcome.
Thank you

JackieBadger 11-10-2012 06:01 PM

without knowing what your library prep is....MiSeq is spec'd to sequence 2x250base. So if your total product including adapters etc is below 500 bases you can sequence your product using paired ends and stitch together the over lapping sequences.
you can in theory sequence any size fragment to to the total possible.

vl80 11-10-2012 06:13 PM

Thank you very much for the reply. I wonder if anyone has attempted this modification before and if there are any publications? I tried to search for it but was not successful.

JackieBadger 11-11-2012 04:22 AM

what modification?
Nextera library prep routinely sequences short fragments.
Just get your PCR product and run it through the TrueSeq library prep and sequence it.
Like I said, there are many ways to prepare a library for sequencing.


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