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JBKri 09-26-2018 02:59 AM

best approach in dealing with low input DNA
 
Hi everybody,
we already posted this in the Nanopore forums, but we wanted to have a better chance of receiving answers so we post it again here.

We are planning in our lab to perform an experiment of de novo whole genome sequencing of bacterial gDNA starting with a extremely limited amount of DNA (about 2-3 ng).
We are considering the SQK-RPB004, Rapid PCR Barcoding Kit to amplify with PCR this initial small amount.

In the knowledge section we found by the way 3 different protocols that use other kits (SQK-LSK108), (SQK-LSK109) (SQK-RAD004) with the idea of avoiding the PCR step including instead a WGA step (whole genome amplification).

Does anybody have some thoughts to share about this?
What is in your opinion the better approach to solve this problem?

Thanks in advance!
Chiara & Jon
HCMR; Institute Of Marine Biology


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