Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2016/07/26
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Major Aims1. 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: Nothing new since 7/24. Two T-75 flasks are growing for René's work and my own.
Update: RNeasy kit ordered. Cells are still growing. Will harvest on Thursday, 7/28.
Update: transformation worked (hundreds of colonies) but plasmid midiprep failed. Maybe picked wrong colony? Trying again tonight + tomorrow. SK-N-SH and K562 cultures expanded from PS-α to PS-β. Still need to check probes.
Update: fluorescence microscopy images taken again of the samples. Cells were harvested by immersion in TRIzol, then stored at -80 C. Will perform RNeasy preps tomorrow in preparation for RNA-seq. Well 1 (U2OS-PcTF +dox, post 96 hr):
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