Hi,
I have an issue I need to resolve regarding multiplexing libraries for a Hiseq run.
There are 96 libraries, I am making them with Truseq V2 RNA prep kits which have 24 available barcodes. So Each barcode will be used 4 times. I have 8 lanes in which to run my libraries. Am I better off:
(i) multiplexing 24 per lane and having essentially 2 technical replicates per library or
(ii) multiplexing 12 per lane and having 1 technical replicate.
My problem is that whilst scheme (i) might reduce impact of variation in number of reads per lane, having more libraries multiplexed may also increase the disparity (variance) between numbers of reads per library.
What other reasons are pro/con each of these schemes? Your thoughts are greatly appreciated. I did try and search the forum and keep an eye out for this as I knew I would be doing this prep for the last few months but haven't seen discussion. Pointers to relevant info also appreciated
I have an issue I need to resolve regarding multiplexing libraries for a Hiseq run.
There are 96 libraries, I am making them with Truseq V2 RNA prep kits which have 24 available barcodes. So Each barcode will be used 4 times. I have 8 lanes in which to run my libraries. Am I better off:
(i) multiplexing 24 per lane and having essentially 2 technical replicates per library or
(ii) multiplexing 12 per lane and having 1 technical replicate.
My problem is that whilst scheme (i) might reduce impact of variation in number of reads per lane, having more libraries multiplexed may also increase the disparity (variance) between numbers of reads per library.
What other reasons are pro/con each of these schemes? Your thoughts are greatly appreciated. I did try and search the forum and keep an eye out for this as I knew I would be doing this prep for the last few months but haven't seen discussion. Pointers to relevant info also appreciated
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