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Mixing old and new kits
Hi, we have one of the old RNA-seq kits from Illumina (the 8 sample kit) and never purchase the multiplexing set. We also have one of the new Set A TruSeq (also RNA-seq) kits that contains the multiplexing (indexing) adaptors. My question is, has anyone used the indexing adaptors from the new kit in place of the standard adaptors from the old kit? Does anyone foresee any issues with this?
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The product which you ligate the adapter to is the same in the new and old library prep kits, an A-tailed, dsDNA molecule so the adapters would ligate. But you would then need to follow up with the PCR primers from the TruSeq kit as well. And make sure you use a cluster generation & sequencing kit compatible with TruSeq libraries. You will be left with a half used original kit, now good for nothing, and a TruSeq kit which is short on adapters and PCR primers.
My question would be "what's the point?". Just use the new TruSeq kit reagents for the entire prep and save your old kit for libraries which don't require multiplexing. -
I agree with "kmcarr" that the new TruSeq adapters should technically work with the reagents from the old library prep kit, however, as far as I know, the adapter as well as the PCR primer concentrations are not the same. For now, Illumina will not release the concentrations of the new TruSeq adapters and primers. If you do end up using the new TruSeq adapters and primers with your old reagents, you may have to adjust the concentrations.Comment
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agreeing as well!
Thanks for the replies. About the time we posted this we sat down and decided it wasn't worth the headache. We'll just run a full lane for each sample using the old kit, relish in twice the data and break out the new one nice and fresh for the next study.Comment
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