What is the minimum quantity of total RNA below which people would not attempt the standard TruSeq mRNA library prep protocol, but would instead use DSN normalization, or an amplification method like SMARTer or the NuGen kit? The current protocol recommends at least 100 ng total RNA, and I've had success making libraries with even less. Is there reason (either theory or data) to believe that less RNA won't work, and if so, why won't it work? If one generates a library using 10 ng total RNA, sees a nice peak on the BioA that quantifies nicely by qPCR, is there some reason the doubt the sequencing results produced from that library?
Any opinions would be very welcome. Thanks!
Any opinions would be very welcome. Thanks!
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