Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2016/08/22

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Overview

Transfected K562 cells on 8/20, following same protocol as before (from 2015-06-03).

K562 KAH132 Results - Microscopy

24hr

Well 1:
2016-08-21 Well 1 KAH132-MV2 phase mCherry 10x.jpg


Well 2:
2016-08-21 Well 2 KAH132-MV2 phase mCherry 10x.jpg


Well 4:
2016-08-21 Well 4 KAH132-MV2 phase mCherry 10x.jpg


Well 5:
2016-08-21 Well 5 KAH132-MV2 phase mCherry 10x.jpg



48 hr

Well 2:
2016-08-22 Well 2 KAH132-MV2 phase mCherry 10x.jpg


Well 5:
2016-08-22 Well 5 KAH132-MV2 phase mCherry 10x.jpg


K562 KAH132 Results - Flow Cytometry

24 hour:
Screen Shot 2016-09-20 at 11.10.45 AM.png


48 hour:
Screen Shot 2016-09-20 at 11.10.50 AM.png


Sample ID % RFP
24hr A 84.8
24hr B 84.9
48hr A 28
48hr B 26

K562 RNeasy prep

Harvested cells today, RNeasy prep. Concentrations/calculations for cDNA synthesis:

Sample ID ng/µL µL for 2 µg RNA (8 µL max) µL H2O qPCR Sample ID
K562 KAH132 24hr 1 917 2.2 5.8 K562_E008
K562 KAH132 24hr 2 837 2.4 5.6 K562_E009
K562 KAH132 48hr 1 879 2.3 5.7 K562_E010
K562 KAH132 48hr 2 638 3.1 4.9 K562_E011

Following steps from cDNA synthesis protocol.