Hi,
I posted this in the library prep forum but I assume it might fit in here better:
I want to use the Illumina TruSEq Kit for SAGE analysis: getting only one read per transcript. To achieve this, I want to fragment the RNA (or prior enriched mRNA) and then perform another rounds of polyA enrichment in order to only get the polyA streches with app. 100-200 bp of UTR region. cDNA synth. will be performed using polyT primers instead of the random ones.
I included a very ugly cartoon of the method as attachment.
Has anybody of you tried a similar protocol, am I missing some pitfalls?
In keen anticipation
Moritz
I posted this in the library prep forum but I assume it might fit in here better:
I want to use the Illumina TruSEq Kit for SAGE analysis: getting only one read per transcript. To achieve this, I want to fragment the RNA (or prior enriched mRNA) and then perform another rounds of polyA enrichment in order to only get the polyA streches with app. 100-200 bp of UTR region. cDNA synth. will be performed using polyT primers instead of the random ones.
I included a very ugly cartoon of the method as attachment.
Has anybody of you tried a similar protocol, am I missing some pitfalls?
In keen anticipation
Moritz
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