Rui:LabNotes/SingleCell/2011-1-27
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Characterization of positive control
*Pure cDNA w/o primer carry-over *1st round of AMpure beads purification, 20ul --> 36ul beads (1.8*) --> 36ul water *2nd round of AMpure beads purification, 30ul --> 21ul beads (0.7* suggested by Alice) --> 36ul water
Concentration Nanodrop [ng/ul] Qubit [ug/ml] 1st Hues54 38.67 (2.16; 2.07) 3.76 Hues6-ES 58.78 (2.11; 2.27) 6.05 Hues6-EB 55.25 (2.07; 2.04) 6.54 2nd Hues54 19.72 (2.36; 2.35) 1.29 Hues6-ES 40.1 (2.19; 2.24) 2.44 Hues6-EB 35.92 (2.21; 2.14) 2.64
File:First round beads.jpg
File:Second round beads.jpg
Titration of Exonuclease I
*Goal: high enough to remove primer/unspecific amplification on NTC; low enough to avoid interference to cDNA stability and following steps *Positive control: diluted cDNA w/o primer *Negative control: Rui water *Additional note: ds DNA treatment for single cell???