Haynes Lab:Notebook/CRISPR Editing/2016/05/30: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</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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'''qPCR for ChIP!'''<br>
'''qPCR for ChIP!'''<br>
Primer sets
Primer sets
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Primer set'''
| align="center" style="background:#f0f0f0;"|'''Target'''
| align="center" style="background:#f0f0f0;"|'''F primer'''
| align="center" style="background:#f0f0f0;"|'''R primer'''
| align="center" style="background:#f0f0f0;"|'''amplicon length'''
|-
| 1||g025||P331 ||P329 ||205
|-
| 2||g031, g034||P308||P160||202
|-
| 3||GAPDH||P203||P204||
|}
<br><br>
'''Plate 1'''<br>
Luc14
:Reps 1-3
::H3K27me3, IgG, input
:::Primer set 1, Primer set 2, Primer set 3
<br>
[[Image:16.05.30 qPCR Plate 1 screen shot.png]]
<br>
Primer master mix for primer sets 1, 2, and 3<br>
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''1 wells'''
| align="center" style="background:#f0f0f0;"|'''34'''
|-
| 2x SYBR mix||7.5||255
|-
| 750nmol F/R primer mix||3||102
|-
| PCR water||2.5||85
|-
| total||13||442
|}
<br><br>
<br><br>
'''Redo nested PCR of the KAH228/g034 experiments'''<br>
Did it with the wrong primers the first time, need to redo with P149/176.
<br><br>
'''Redo nested PCR of the Luc14 g034 samples'''<br>
Did it with the wrong primers the first time, need to redo with P149/176.




Latest revision as of 01:49, 27 September 2017

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05/30/2016

qPCR for ChIP!
Primer sets

Primer set Target F primer R primer amplicon length
1 g025 P331 P329 205
2 g031, g034 P308 P160 202
3 GAPDH P203 P204



Plate 1
Luc14

Reps 1-3
H3K27me3, IgG, input
Primer set 1, Primer set 2, Primer set 3



Primer master mix for primer sets 1, 2, and 3

' 1 wells 34
2x SYBR mix 7.5 255
750nmol F/R primer mix 3 102
PCR water 2.5 85
total 13 442






Redo nested PCR of the KAH228/g034 experiments
Did it with the wrong primers the first time, need to redo with P149/176.



Redo nested PCR of the Luc14 g034 samples
Did it with the wrong primers the first time, need to redo with P149/176.




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