User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/30: Difference between revisions
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* | * K562 transduction (trial 2) | ||
* Cell culture: passaged U2OS 1:10, K562 1:10 | |||
* Cell culture: started Luc14 & Gal4EED/luc cultures (4 mL 20% FBS/ 1% P/S) | |||
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''' | '''K562 transduction (trial 2)'''<br> | ||
* See ##/##/## for transgene/virus list | |||
* Cell stock passaged 1:10 on 9/22/14. Assume 1.0E6 cells/mL | |||
* Added 1 mL complete culture medium to 6x 15 mL conicals | |||
* Added 80 μL cells to each tube | |||
* LaBaer lab | |||
** Bihazard set-up: Wescodyne waste and rinse; 10 mL in 2x 50 mL conicals | |||
** Added 5.0 μL 2.4 mg/mL polybrene (thawed) to each cell sample. Swirled to mix. | |||
** Added 400 μL virus (thawed/ spun/ pipetted to remix) to each cell sample. Swirled to mix. (rinsed pipette tip before discarding) | |||
***Note: had some left-over virus; marked these with black dot on caps, placed back into the -80°C box | |||
** Spun at 2250 rpmn (1178 rcf) for 25 min. | |||
** Pipetted u/d to resuspend, transferred cells to 6 mL plate | |||
** Grew at 37°C. Will check GFP expression tomorrow. | |||
Revision as of 14:50, 30 September 2014
 Pc-TF Genomics
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09/30/14
K562 transduction (trial 2)
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