User:Karmella Haynes/Notebook/PcTF Genomics/2014/11/15: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</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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== | ==11/15/14== | ||
<!-- Precede finished items with a checkmark ✓ --> | <!-- Precede finished items with a checkmark ✓ --> | ||
* | * Transfection: KAH126/MV2 into U2OS | ||
---- | ---- | ||
''' | '''Transfection: KAH126/MV2 into U2OS'''<br> | ||
Strategy: instead of trying to increase transfection efficiency, do a "quasi-subcloning" where transfected cells are separated into smaller wells, and wells showing highest RFP will be selected, pooled, and re-plated. | |||
* 6-well plate seeded on 11/14/14 (~2.5E5 cells/well) | |||
* Change medium to p/s-free before transfection (4 mL per well) | |||
* Use standard Lipofectamine LTX procedure w/ 2000 ng plasmid | |||
# KAH126/MV2 | |||
# mock | |||
# no transfection | |||
* Grow overnight, look for RFP next day | |||
* Next day: harvest transfected cells and use 1/2 (5.0E5) for 2 different plates | |||
** 24-well plate - ~2.1E4 cells/well | |||
** 96-well plate - ~5.2E4 cells/well | |||
Latest revision as of 00:31, 27 September 2017
Pc-TF Genomics | Main project page Previous entry Next entry |
11/15/14
Transfection: KAH126/MV2 into U2OS
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