Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/04/23: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Neon transfection trial on K562 & KAH126-MV2</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Neon transfection trial on K562 & KAH126-MV2</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 00:55, 27 September 2017
Neon transfection trial on K562 & KAH126-MV2 | Main project page Previous entry Next entry |
OverviewThe neon transfection system is an electroporation instrument that allows you to transfect very small quantities of mammalian cells inside a conductive 10µL pipette tip. We were given a demo to try out in our lab. This is my attempt using K562 cells and the KAH126-MV2 plasmid. NotesI followed the procedure as outlined in the neon transfection kit. K562 parameters can be found here. K562 culture density before resuspending in neon transfection buffer: 1.5E6 cells/mL. Cells were resuspended to 2E7 cells/mL in 60µL total. KAH126-MV2 plasmid concentration is 809 ng/µL. Volume of a.b.-free medium in each well of the 6-well plate: 2 mL. For each well, I added 10µL of the resuspended cells to 1µL of plasmid or water in a sterile microcentrifuge tube. Well layout:
The 6-well plate was then placed in the 37°C incubator. Will check fluorescence tomorrow for an estimate of efficiency.
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