Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/06/10

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K562 Transfection Results & RNA Miniprep Main project page
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Overview

Day 2 of collecting flow cytometry data, fluorescence microscopy data, and harvesting cells for RNA.

Also need to set up two more samples for a "no plasmid" control transfection. Current cell density of K562 culture is 420,000 cells/mL. Need to wait until culture is around 800,000 cells/mL before proceeding with mock transfection.

Following the protocol for RNA miniprep found here.

Fluorescence Microscopy

Well 4: pMaxGFP, post-48hr

Well 5: KAH126-MV2, post-48hr

Well 6: KAH126-MV2, post-48hr

Flow Cytometry

Plot 1 (FSC-A/SSC-A)Plot 4 (FL3-A)Plot 6 (FL1-A)
AllCellsM1M2
CountVolume (µL)Events / µLCountEvents / µL% of AllCount% of This PlotCount% of This Plot
A02 0 untreated3,967103972,93029373.86%130.44%150.51%
A06 4 pMaxGFP21,81410218110,666106748.90%6,37859.80%9,94593.24%
A07 5 KAH126-MV211,1551011165,54755549.73%67412.15%3686.63%
A08 6 KAH126-MV28,363108363,63636443.48%3309.08%240.66%
A09 0 untreated3,881103882,86228673.74%20.07%10.03%

RNeasy / Trizol

Samples are stored at -80 °C.



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