Blue:RNA-Seq Experiments:06122013
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Library:
Purpose: Optimize T20/TSO and library amplification. Tested UHRR (250pg) for: (1) Betaine; (2) Conc of T20 primer; (3) one-step versus two-step T20/TSO.
Outcome: Variability in amplification possibly due to limiting template. Betaine seemed to enhance efficiency. Increasing T20 conc had no effect. Combining T20 and TSO in one step (beads purification prior to PCR) increased efficiency but also led to high primer dimers.