Hi All,
being new here and to single-cell RNA-Seq too, I have several questions on the choice of amplification techniques, but haven't found a thread that would deal with this question.
It seems to me that there are two major approaches to RNA amplification, PCR-based (e.g. SmartSeq) and in vitro transcription based. I'm interested in first-hand experience on which one the more experienced recommend for RNA-Seq from 20-200 cells when we are interested in differential expression between different tissue types? Low expressor genes are of interest, as would potentially isoforms be, but the latter is secondary to as accurately counting transcripts as possible.
I would appreciate advice on this or pointing me to places where this has been discussed already (sorry for re-post if so). Thanks!
being new here and to single-cell RNA-Seq too, I have several questions on the choice of amplification techniques, but haven't found a thread that would deal with this question.
It seems to me that there are two major approaches to RNA amplification, PCR-based (e.g. SmartSeq) and in vitro transcription based. I'm interested in first-hand experience on which one the more experienced recommend for RNA-Seq from 20-200 cells when we are interested in differential expression between different tissue types? Low expressor genes are of interest, as would potentially isoforms be, but the latter is secondary to as accurately counting transcripts as possible.
I would appreciate advice on this or pointing me to places where this has been discussed already (sorry for re-post if so). Thanks!