Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2016/07/26: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Today's project is...</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Today's project is...</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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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. | 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):<br> | |||
[[Image:2016-07-26_Well_1_1_ug-mL_doxycyclin_phase_mCherry_10x.jpg | 250px]] | |||
Well 2 (U2OS-PcTF +dox, post 96 hr):<br> | |||
[[Image:2016-07-26_Well_2_1_ug-mL_doxycyclin_phase_mCherry_10x.jpg | 250px]] | |||
Well 3 (U2OS-PcTF +dox, post 96 hr):<br> | |||
[[Image:2016-07-26_Well_3_1_ug-mL_doxycyclin_phase_mCherry_10x.jpg | 250px]] | |||
Well 1 (U2OS ΔTF (transient), post 96 hr):<br> | |||
[[Image:2016-07-26_Well_1_KAH132-MV2_phase_mCherry_10x.jpg | 250px]] | |||
Well 2 (U2OS ΔTF (transient), post 96 hr): <br> | |||
[[Image:2016-07-26_Well_2_KAH132-MV2_phase_mCherry_10x.jpg | 250px]] | |||
Well 3 (U2OS ΔTF (transient), post 96 hr): <br> | |||
[[Image:2016-07-26_Well_3_KAH132-MV2_phase_mCherry_10x.jpg | 250px]] | |||
No photos taken of U2OS-PcTF without dox treatment. | |||
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Latest revision as of 01:50, 27 September 2017
Today's project is... | Main project page Previous entry Next entry |
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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