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

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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[[Image:2015-04-15_KAH187-MV8_digest_annotated.png]]
[[Image: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.


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Latest revision as of 00:54, 27 September 2017

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

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.