Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/01/29: Difference between revisions
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Gel image from plasmids purified over the last three weeks: | Gel image from plasmids purified over the last three weeks: | ||
[[Image:2015-01-29_Ben_KAH126-MV2_digestion.JPG | 500px | EcoRI & PstI digestion of KAH126-MV2 plasmid]] | |||
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. | |||
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Revision as of 21:06, 29 January 2015
 RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines
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SummaryToday I'll be performing another transfection of SKNSH samples, as well as purifying plasmid and running it on a gel to check for purity.
Goals
ProtocolPlasmid mini-prep: Follow instructions in kit transfection with lipofectamine Restriction digestion setup:
Incubate for 30min at 37°C, then run on a 1% agarose gel with 1kb ladder.
NotesObservations, measurements, placeholder info etc. go here.
Results/ConclusionsPlasmid concentration:
Gel image from plasmids purified over the last three weeks:
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.
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