Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2016/07/24
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1. U2OS-PcTF and U2OS-TF stable cell lines ChIP-PCR on (1) PcTF, (2) H3K27me3, (3) H3K4me3, and (4) PolII before, during, and after PcTF expression at endogenous enhancer(s)
Update: Dr. Haynes has ordered primers (check this). Cell lines are being split into plates by Rene and Ben. Looks like the U2OS-TF line is not growing.
To do: Check U2OS-PcTF cell line and split into T-75 for Rene.
Update: KAH126-U2OS cells were split into two 12-well plates. Dox induction began at 5pm today. Will harvest cells for flow cytometry, RNA extraction and cDNA synthesis on 7/28.
Order more of the RNA extraction kit.
Dox induction plan:
Update: don't know status of U2OS cells, need to check on that. Also need to make more KAH132_MV2 plasmid. Transforming DH5α-T cells overnight, will start liquid culture tomorrow afternoon.
Check for levels of probes, order more if needed.
Update: KAH126-U2OS cells were induced on Friday, and U-2OS cells were transfected with KAH132 on Friday. Visible expression in both plates (see below).
KAH126-U2OS, 48 hours post induction.
Will harvest two KAH126 +dox wells, two KAH126 no dox wells, and two U-2OS + KAH132-MV2 transfection wells on 7/26. Will also take photos prior to harvesting for RNA-seq.