Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/02/17

RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines

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Summary In paragraph format, state what you are going to do today so a reader unfamiliar with your project could understand what is going on. Goals ✓ Transfect K562 cells using nucleofector method; transfect with KAH126/MV2 plasmid Protocol example protocol Notes Observations, measurements, placeholder info etc. go here. Results/Conclusions Culture was at 300,000 cells/mL before transfection. Entire culture (9mL, 2,700,000 cells total) was spun down and resuspended into 400uL of nucleofection buffer, then aliquoted 100uL into each of 4 reaction tubes (675,000 cells per reaction). Sample list: Well 1: positive control - nucleofector GFP plasmid Well 2: negative control - no plasmid Well 3: replicate 1 - KAH126/MV2 (PcTF-RFP) Well 4: replicate 2 - KAH126/MV2 (PcTF-RFP) After nucleofection, cells were resuspended in 1mL of medium and then transferred into 4mL total in a 6-well plate.