Shreffler:Notebook/retinoic acid trans signaling/2009/07/28

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Neisha A. Rivers

  • Transfection

Cell count:

Jurkats = 9.20e5 cells 2 conditions (100nM RA and DMSO): 4.5e5 cells per condition

3T8 = 1.39e6 cells 2 conditions (100nM RA and DMSO) : 4.5e5 cells per condition


Addition of Plasmids:

RARE = 2 ug /189.7 ug/ml = 10.5 ul RARE

Pcmv = 2 ug /744 ug/ml = 2.6 ul Pcmv

RXR - 2 concentrations

* 0.1 ug / 3 ug/ml = 0.03 ul RXR
* 0.4 ug/ 3 ug/ml = 0.13 ul RXR

RAR - 2 concentrations

* 0.1 ug / 1.1 ug/ml = 0.09 ul RAR
* 0.4 ug/ 1.1 ug/ml = 0.36 ul RAR 

Incubation overnight