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Old 02-04-2011, 08:03 AM   #18
TonyBrooks
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Location: London

Join Date: Jun 2009
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Quote:
Originally Posted by li zhang View Post
Hi, Im a beginner at sequencing. I have several might silly questions. Are RNA-seq and transcritome sequencing the same thing and using the same protocol? Can paired-end squencing be adopted in transcritome sequencing? If so what is the protocol? Much appreciate!
RNA-seq and transcriptome sequencing are essentially the same protocol, but with different aims.
With RNA-seq, the aim is to estimate transcript/exon, and hence expression, levels. For this, you typically require lots of reads in order to get sufficient sensitivity.
The aim of transcriptome sequencing is to sequence the transcriptome - i.e. you're looking for polymorphisms in sequence, depth is much less of an issue.
Of course, you can "do" transcriptome sequencing with RNA-seq data, but not necessarily vice versa.

With regards rRNA sequencing, we have always used PolyA+ selection. Our experience with ribominus is that it doesn't remove sufficiently enough RNA for sequencing.
We're also just about to sequence a DSN treated library. We've been told that this method has excellent results for rRNA removal. It'll be interesting to see the data.
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