Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/05/21

DNA Concentration of pMaxGFP

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Overview The sample left to dry overnight has evaporated down to a volume of under 30 µL. I'm taking another 500 µL of the plasmid and performing a sodium acetate-ethanol precipitation, following the instructions in the Sigma GenElute Plasmid Maxiprep Kit from yesterday. Final volume of the precipitation concentration will be 100 µL, a theoretical 5-fold increase in concentration. DNA Quantification I ran each sample in duplicate, and at straight (undiluted) and 10x diluted concentrations. Sample ID Conc. (ng/µL) St Dev (ng/µL) 260/280 pMaxGFP pre-concentration 486.333 7.966264997 1.888 evap conc. 2090.9 17.05 1.90 evap conc, 10x dil 190.1 1.86 1.91 precip conc. 2297.7 43.69 1.90 precip conc, 10x dil 205.7 5.06 1.82 There's a little bit of saturation of the instrument at concentrations around 2000 ng/µL, but overall the results look very consistent with each other and with expected values. Recovery from the precipitation is 85%.