Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/05/19
|Setup for K562 transfection optimization||Main project page|
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K562 cells are in PS-α right now. Once they're ready for expansion into PS-β, I'll be setting aside a fraction to grow in antibiotic-free medium for a transfection optimization experiment. Here's the general layout of the transfection procedure I'll be performing.
1. Prepare master mixes of Opti-MEM, DNA, & PLUS Reagent.
2. Prepare master mixes of Opti-MEM and LTX Reagent.
3. Incubate at room temperature for 5 minutes, then combine the two master mixes for each sample (100+µL total for each sample).
4. Take cells out of the incubator and add 50 µL of mixed DNA + LTX into each well (duplicates for each sample).