Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/01/26

RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines

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Summary

Still checking on the cultures, going to harvest the ones induced to create PcTF-RFP later in the week if their expression levels look good. Also submitting the RNA harvested last week to DNASU, where they'll create cDNA and then quantify it using real-time PCR.

Goals

Monitor cultures (SKNSH, U2OS, & KAH126-U2OS)
Photograph fluorescence of transfected/induced cultures
Harvest cultures that are on day 4 of transfection/induction
Quantify RNA prior to submission to DNASU lab for sequencing

Protocol

example protocol

Notes

Observations, measurements, placeholder info etc. go here.

Results/Conclusions

Culture Status

SKNSH (6-well plate): 25% confluence, replace with fresh medium
SKNSH (T-75): 50% confluence, replace with fresh medium
KAH126-U2OS (T-75): 80% confluence, replace with fresh medium
U2OS (T-75): 100% confluence, passage

RNA Quantification

	Replicate 1	Replicate 2	Measurement
Well 1 - dox	0.95	0.577	260
	0.475	0.276	280
	2	2.089	260/280
	760.119	461.95	ng/µL
Well 2 - dox	0.661	0.647	260
	0.313	0.308	280
	2.11	2.101	260/280
	528.495	517.669	ng/µL
Well 4 - negative control	0.908	0.457	260
	0.443	0.224	280
	2.048	2.037	260/280
	726.307	365.656	ng/µL