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

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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.  
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%.


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Revision as of 15:41, 21 May 2015

DNA Concentration of pMaxGFP <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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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. 2090.8875 17.05470846 1.904
evap conc, 10x dil 190.106 1.863933475 1.907
precip conc. 2297.7305 43.69000669 1.901
precip conc, 10x dil 205.733 5.06147034 1.817

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%.