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

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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)

2015-06-09 Well 1 pMaxGFP phase GFP 10x.jpg

Well 2: KAH126-MV2 (post 24-hr sample, rep 1)

2015-06-09 Well 2 KAH126-MV2 phase mCherry 10x.jpg

Well 3: KAH126-MV2 (post 24-hr sample, rep 2)

2015-06-09 Well 3 KAH126-MV2 phase mCherry 10x.jpg

Well 4: GFP (post 48-hr sample)

2015-06-09 Well 4 pMaxGFP phase GFP 10x.jpg

Well 5: KAH126-MV2 (post 48-hr sample, rep 1)

2015-06-09 Well 5 KAH126-MV2 phase mCherry 10x.jpg

Well 6: KAH126-MV2 (post 48-hr sample, rep 2)

2015-06-09 Well 6 KAH126-MV2 phase mCherry 10x.jpg

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.