Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/05/20: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Plasmid Maxiprep of pMaxGFP</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Plasmid Maxiprep of pMaxGFP</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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Latest revision as of 00:58, 27 September 2017
Plasmid Maxiprep of pMaxGFP | Main project page Previous entry Next entry | ||||||||
OverviewI'm making a lot of the pMaxGFP plasmid for use in transfection (Neon transfection, lipofectamine, etc.), as these processes require a very high concentration of very pure DNA. ProcedureYesterday I set up a 150mL culture of LB-Ampicillin from a DH5α-Turbo colony transformed with the pMaxGFP plasmid. I spun down the culture at 5000g for 10 minutes and discarded the supernatant, then followed the instructions for plasmid prep as laid out in the Sigma GenElute HP Endotoxin-Free Plasmid Maxiprep Kit. Plasmid concentration was measured on the Take3 Nanodrop. DNA QuantificationFinal volume of the purified plasmid is approximately 1.5 mL.
I took 100µL of the purified plasmid and transferred it to a new sterile tube and am leaving that tube open in the BSC overnight to evaporate off and concentrate. Tomorrow I'll also perform a sodium acetate-ethanol precipitation with another aliquot of the plasmid.
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