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Old 01-31-2010, 08:27 PM   #1
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Default On flowcell, amplification-free RNA-seq

(I like my title better :-)

Nat Methods. 2010 Feb;7(2):130-2. Epub 2010 Jan 17.
FRT-seq: amplification-free, strand-specific transcriptome sequencing.
Mamanova L, Andrews RM, James KD, Sheridan EM, Ellis PD, Langford CF, Ost TW, Collins JE, Turner DJ.

[1] The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK. [2] These authors contributed equally to this work.
We report an alternative approach to transcriptome sequencing for the Illumina Genome Analyzer, in which the reverse transcription reaction takes place on the flowcell. No amplification is performed during the library preparation, so PCR biases and duplicates are avoided, and because the template is poly(A)(+) RNA rather than cDNA, the resulting sequences are necessarily strand-specific. The method is compatible with paired- or single-end sequencing.

PMID: 20081834
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Old 01-31-2010, 10:39 PM   #2
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Wow. I know what I'm reading tomorrow morning.
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Old 02-04-2010, 05:46 PM   #3
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I want to try it now.
Someone has tried this progression method ?
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Old 06-03-2010, 08:41 AM   #4
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that sounds incredible
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Old 06-08-2010, 01:40 PM   #5
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Gary Schroth appeared to be upset at the quality of the "normal library prep' RNA-Seq they did for this paper at the Midwest NGS Users Meeting.
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Old 06-08-2010, 06:28 PM   #6
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If this paper has any verisimilitude, then that the 'standard mRNA-seq' library prep is quantitative is dubious. Perhaps we might have marched upon the dubiousness sooner, were it not for the clutter of papers lauding the 'standard mRNA-seq's technical reproducibility using statistics designed to assess association instead of agreement.
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