Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/01/29
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Today I'll be performing another transfection of SKNSH samples, as well as purifying plasmid and running it on a gel to check for purity.
Plasmid mini-prep: Follow instructions in kit
For reference on plasmid restriction digestion and expected size, see Karmella's notes here. Restriction digestion setup:
Incubate for 30min at 37°C, then run on a 1% agarose gel with 1kb ladder.
Observations, measurements, placeholder info etc. go here.
Gel image from plasmids purified over the last three weeks:
Expected size fragments are 4.5kb for MV2 backbone, 1.9kb for PcTF-RFP insert
Samples from left to right:
Ladder, 31ng/μL, 38ng/μL, empty, 61ng/μL, empty, 66ng/μL, 378ng/μL, ladder, negative control
All plasmids collected look fairly clean, though only one is of high enough concentration to use in transfection.
SKNSH cells were transfected and placed back in the incubator. Tomorrow, check for transfection efficiency, and if >25% harvest on Monday.