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Performed TotoRNAseq protocol (RT-based) on mESC single cell sorted 96-well plate (Exp 03032013) Repeated TSO and PCR on precipitate from 02272013 and 02282013 Results of smarter PCR: [[File:03032013_smarterPCR1.jpg]] Appears that the undissolved precipitate contained the majority of the library. Note that the RnaseIII sample still had a significant amount of undissolved precipitate... this may have resulted in the lower efficiency For the mESC sample, there was a large pellet, but I vortexed the pellet with 70% ethanol much longer to remove the extra salt The fragmentation time for lane 1 was 2 minutes and lane 3 was 3 minutes. Maybe 4 minutes would show a better size range...
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