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Old 05-08-2014, 05:39 AM   #4
Senior Member
Location: New England

Join Date: Jun 2012
Posts: 199

The reason for using the 2-step PCR for amplicon sequencing is so that you can use one set of primers to produce all your amplicons, and the Nextera dual-index primers to index/add the rest of the adapter. This cuts down on cost dramatically. It would be very expensive to order a different set of primers for every indexed sample.
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