Quantification of mRNA

[daisieh]

Hi all,

This may be a stupid question, but I was wondering if anyone knew what sort of mRNA recovery rate one generally expects from the Sera-Mag beads that come in the Illumina mRNA-seq library kit. I'm getting really low recovery rates, on the order of 1% of the original transcript present, but I don't know if that's normal or not.

[pmiguel]

Are you talking about mRNA recovery from total RNA? If so, 1% is very good in my opinion. Except for extremely message-rich tissues (eg, fish gonads) I rarely see poly A RNA yields above 1%. Especially after 2 cycles of enrichment. (Which is what the TruSeq RNA prep kit uses.)

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Phillip