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We want to sequence miRNA from 42 samples and are wondering about the best strategy for pooling and multiplexing. Has anyone here experience in multiplexing such a large number of samples in one lane?
We will run this on SOLiD, but are interested in input from Illumina users as well.
We plan to use 3 lanes, to get 7-8 M reads per sample. We can either pool all 42 libraries and split the pool on three lanes, OR we can make 3 sub-pools, each with 14 libraries, and run on separate lanes.
3 sub pools is laborious and expensive, since we need to run 3 separate emulsion PCRs. Our concern is however, how well we would be able to pool all 42 libraries and get equal representation from each?
We will run this on SOLiD, but are interested in input from Illumina users as well.
We plan to use 3 lanes, to get 7-8 M reads per sample. We can either pool all 42 libraries and split the pool on three lanes, OR we can make 3 sub-pools, each with 14 libraries, and run on separate lanes.
3 sub pools is laborious and expensive, since we need to run 3 separate emulsion PCRs. Our concern is however, how well we would be able to pool all 42 libraries and get equal representation from each?
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