Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/04/15

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.pngRNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Plasmid Maxiprep of KAH187-MV8

--Notes-- During the plasmid purification procedure, I accidentally mixed the lysate flowthrough (containing plasmid) with the column preparation solution that was run through the Binding Column. Proceeding as normal from this point forward. Hopefully doesn't influence yield.


--DNA Quantification--

Measured using the Take3 plate:

Sample ID Concentration (ng/µL) St Dev (ng/µL)
KAH187-MV8 163.5 7.1

--Restriction Enzyme Digest--

Reagent Volume
DNA 600ng (4uL)
10x FastDigest Buffer 3
EcoRI 1
ApaLI 1
Water 21
Total 30

Digest for 15 minutes at 37°C.


--Gel Electrophoresis--

2015-04-15 KAH187-MV8 digest annotated.png

Expected size fragments from EcoRI/ApaLI digestion: 3951, 1243, 1135, 288, 234.

Gel was run a little too long, ~300bp fragment sizes are difficult to visualize.