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Old 07-28-2017, 07:26 AM   #1
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Location: Norman, OK

Join Date: Apr 2017
Posts: 4
Default Questions about RADseq oligo

I will be genotpying ~500 samples using the original RADseq protocol, and I have a couple of questions.

First, is desalting a sufficient purification method for the barcoded adapters? I plan to order P1 adapters with 48 different barcodes, and other purification methods add significantly to the cost so I'm wondering if the added cost is worth it.

Second, I would like to use combinatorial multiplexing by using barcodes for individual samples plus an index to identify separate pools of samples. In ddRAD, the index is incorporated into one of the PCR primers, but in the original RAD protocol the PCR primer is shorter and has no appropriate place for an index.

Does anyone have any suggestions for how to modify the original RADseq protocol to allow combinatorial multiplexing?
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