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Old 10-26-2018, 04:26 AM   #1
rjlebbink
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Location: Europe

Join Date: Apr 2014
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Default Minion sequencing

Hi all,

I have a very basic question. We have 10 samples that we would like to sequence using MinION. These are PCR amplicons that we would like to multiplex in a single run.

Is it possible for me to attach index/adapters to these amplicons by a second PCR using custom primers that I can order as a standard primer from any primer company? If so, are there any recommendations for this? Can I just add the index right upstream of the gene-specific primers, or do I need to add an additional adapter up/downstream to allow sequencing of the amplicons via MinION? I am just trying to understand whether I need to use the nanopore PCR Barcoding Kit, or whether I can just add these (for much less $$ and effort) by PCR myself?

Cheers
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