Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2016/08/16

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Overview

Repeated dox dose curve with 6-well plates (see previous days). Samples are 1.1-1.3 (1 µg/mL dox), 6.1-6.3 (31 ng/mL dox), 7.1-7.3 (16 ng/mL dox), and 8.1-8.3 (no dox).

Harvested cells with TRIzol and extracted RNA using RNeasy mini kit. Concentrations/calculations for cDNA synthesis:

Sample ID ng/µL µL for 2 µg RNA (8 µL max) qPCR Sample ID
1.1 150 8 U2OS_E019
1.2 197 8 U2OS_E020
1.3 180 8 U2OS_E021
6.1 241 8 U2OS_E022
6.2 221 8 U2OS_E023
6.3 195 8 U2OS_E024
7.1 173 8 U2OS_E025
7.2 207 8 U2OS_E026
7.3 227 8 U2OS_E027
8.1 265 7.5 U2OS_E028
8.2 345 5.8 U2OS_E029
8.3 277 7.2 U2OS_E030

Following steps from cDNA synthesis protocol.