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

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==Plasmid Maxiprep of KAH187-MV8==
==Plasmid Maxiprep of KAH187-MV8==


<b>--Notes--</b>
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.
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.


-----
<b>--DNA Quantification--</b>
Measured using the Take3 plate:
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Sample ID'''
| align="center" style="background:#f0f0f0;"|'''Concentration (ng/µL)'''
| align="center" style="background:#f0f0f0;"|'''St Dev (ng/µL)'''
|-
| KAH187-MV8||163.5||7.1
|}
-----
<b>Restriction Enzyme Digest--</b>
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Reagent'''
| align="center" style="background:#f0f0f0;"|'''Volume'''
|-
| DNA||600ng (4uL)
|-
| 10x FastDigest Buffer||3
|-
| EcoRI||1
|-
| ApaLI||1
|-
| Water||21
|-
| Total||30
|}
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__NOTOC__
__NOTOC__

Revision as of 15:43, 15 April 2015

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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


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