Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/06/09

K562 Transfection Results & RNA Miniprep

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Overview Took fluorescence microscopy images & flow cytometry measurements of all 6 samples today. Harvesting the first three samples for RNAseq. Following the protocol for RNA miniprep found here. Fluorescence Microscopy Well 1: GFP (post 24-hr sample) Well 2: KAH126-MV2 (post 24-hr sample, rep 1) Well 3: KAH126-MV2 (post 24-hr sample, rep 2) Well 4: GFP (post 48-hr sample) Well 5: KAH126-MV2 (post 48-hr sample, rep 1) Well 6: KAH126-MV2 (post 48-hr sample, rep 2) Flow Cytometry Plot 1 (FSC-A/SSC-A) Plot 4 (RED) Plot 6 (GRN) All Cells M1 M2 Sample ID Count Volume (µL) Events / µL Count Events / µL % of All Count % of This Plot Count % of This Plot untreated 2,261 10 226 1,624 162 71.83% 8 0.49% 9 0.55% Well 1 pMaxGFP 12,421 10 1242 3,798 380 30.58% 2,704 71.20% 3,603 94.87% Well 2 KAH126-MV2 5,625 10 563 862 86 15.32% 265 30.74% 70 8.12% Well 3 KAH126-MV2 6,151 10 615 1,149 115 18.68% 298 25.94% 5 0.44% Well 4 pMaxGFP 12,612 10 1261 4,669 467 37.02% 3,178 68.07% 4,372 93.64% Well 5 KAH126-MV2 6,406 10 641 1,057 106 16.50% 293 27.72% 33 3.12% Well 6 KAH126-MV2 6,972 10 697 1,790 179 25.67% 402 22.46% 5 0.28% untreated 1,507 10 151 839 84 55.67% 5 0.60% 1 0.12% RNA Miniprep Follow the instructions given here. Samples are stored at -80 °C.