Greetings everyone,
I am a graduate student making his first foray into NGS. Briefly, I am employing a ddRAD protocol to generate my first NGS dataset. I'll be using Illumina MiSeq.
I would ideally like to include ~48 individuals on a single Illumina MiSeq lane, each with a unique individual barcode/ID.
I understand that performing 48 separate size-selections would probably be ill-advised and may lead to substantial drop-out at certain loci and a loss of data.
Is it possible to anneal my Illumina adapters with unique barcodes to each of the 48 individuals separately, and then pool them in equimolar concentrations to perform a single size-selection, either with a Pippin Prep or with SPRI beads? Is this at all advisable?
Many thanks!
I am a graduate student making his first foray into NGS. Briefly, I am employing a ddRAD protocol to generate my first NGS dataset. I'll be using Illumina MiSeq.
I would ideally like to include ~48 individuals on a single Illumina MiSeq lane, each with a unique individual barcode/ID.
I understand that performing 48 separate size-selections would probably be ill-advised and may lead to substantial drop-out at certain loci and a loss of data.
Is it possible to anneal my Illumina adapters with unique barcodes to each of the 48 individuals separately, and then pool them in equimolar concentrations to perform a single size-selection, either with a Pippin Prep or with SPRI beads? Is this at all advisable?
Many thanks!
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