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I'm using unique dual index primer pairs for my experiments. I'm multiplexing around 12 samples per NextSeq run. I don't know how to create a custom library in BaseSpace that will allow me to assign the unique primer pairs to all my samples as it's limited by a 12 x 8 format (12- i7 indexes and 8-i5 indexes, I need to be able to enter 12 i5 indexes). Any help would be greatly appreciated.
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I ran into a similar issue recently. If you have the Dx version of the NextSeq, there is a software update that can do FASTQ generation on the machine similar to the MiSeq.
The other option is a work around where you will have to create two custom library kit. Each kit can do 8 UDI samples if you set your samples in a diagonal so that each row and column will have an unique index. After that just pool them together in the pool tab in BaseSpace. -
I don't have the Dx version but your workaround will be just fine, I didn't think it would let me combine samples from two different "kits". Thanks for the help!Comment
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custom library
Here's the bulletin from Illumina describing how to create a custom library prep kit in BaseSpace.Comment
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Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.07-01-2026, 11:43 AM
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