Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2016/07/24: Difference between revisions
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<b>1. U2OS-PcTF and U2OS-TF stable cell lines <u>ChIP-PCR</u> on (1) PcTF, (2) H3K27me3, (3) H3K4me3, and (4) PolII before, during, and after PcTF expression at endogenous enhancer(s)</b> | <b>1. U2OS-PcTF and U2OS-TF stable cell lines <u>ChIP-PCR</u> on (1) PcTF, (2) H3K27me3, (3) H3K4me3, and (4) PolII before, during, and after PcTF expression at endogenous enhancer(s)</b> | ||
Update: Cell lines are being split into plates by | 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. | |||
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<b>3. <u>qRT-PCR</u> experiments with TF control plasmid. Cannot select stables because PcTF is toxic. Try normalizing to PcTF levels. Include CDKN2A.</b> | <b>3. <u>qRT-PCR</u> experiments with TF control plasmid. Cannot select stables because PcTF is toxic. Try normalizing to PcTF levels. Include CDKN2A.</b> | ||
Update: | Update: don't know status of U2OS cells, need to check on that. Also need to make more KAH132_MV2 plasmid from transformation plate. Will start liquid culture this afternoon. | ||
Revision as of 11:26, 25 July 2016
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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: 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.
Update: don't know status of U2OS cells, need to check on that. Also need to make more KAH132_MV2 plasmid from transformation plate. Will start liquid culture this afternoon.
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.
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