Haynes Lab:Notebook/CRISPR Editing/2015/03/04: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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|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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==03/04/2015==
==03/04/2015==
Plated cells for transfections tomorrow. Gal4-EED+dox cells plated at 0.16x10^6 cells/well in 12 well plates. Luc14 cells plated at 0.1x10^6 cells / well in 12 well plates. Two plates for each cell type for a total of 4 12-well plates.
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| align="center" style="background:#f0f0f0;"|'''Cell type'''
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<br><br>
Diluted untreated and g046 column purified P151/158 PCR products down to 30ng/ul each.<br>
Cut 250ng DNA with HindIII, 20 minutes at 37deg C and 20 min at 80deg C to inactivate. Did in new PCR machine. <br>
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''DNA'''
| align="center" style="background:#f0f0f0;"|'''250ng'''
|-
| enzyme||0.5
|-
| 10xbuffer||2
|-
| water||9.17
|-
| Total||20
|}
<br>
Dilute 1ul into 20ul water. run 1ul on bioanalyzer.
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__NOTOC__
__NOTOC__

Latest revision as of 00:48, 27 September 2017

Project name Main project page
Previous entry      Next entry

03/04/2015

Plated cells for transfections tomorrow. Gal4-EED+dox cells plated at 0.16x10^6 cells/well in 12 well plates. Luc14 cells plated at 0.1x10^6 cells / well in 12 well plates. Two plates for each cell type for a total of 4 12-well plates.

Cell type treatment rep 260 280 260/280 ng/µL
Luc14 g023 1 0.089 0.048 1.862 89.331
Luc14 g023 2 0.087 0.046 1.909 87.359
Luc14 g023 3 0.088 0.048 1.834 88.229
Gal4-EED +dox g023 1 0.091 0.049 1.869 91.083
Gal4-EED +dox g023 2 0.091 0.05 1.833 91.397
Gal4-EED +dox g023 3 0.096 0.052 1.832 95.767
Luc14 g045 1 0.078 0.043 1.808 77.703
Luc14 g045 2 0.029 0.016 1.75 28.731
Luc14 g045 3 0.067 0.036 1.861 66.736
Gal4-EED +dox g045 1 0.085 0.046 1.836 85.215
Gal4-EED +dox g045 2 0.073 0.041 1.788 73.213
Gal4-EED +dox g045 3 0.067 0.037 1.805 67.248
Luc14 g048 1 0.056 0.03 1.849 56.099
Luc14 g048 2 0.056 0.031 1.795 56.201
Luc14 g048 3 0.06 0.034 1.782 59.902
Gal4-EED +dox g048 1 0.076 0.041 1.844 76.457
Gal4-EED +dox g048 2 0.072 0.039 1.854 71.611
Gal4-EED +dox g048 3 0.077 0.042 1.838 77.038



Diluted untreated and g046 column purified P151/158 PCR products down to 30ng/ul each.
Cut 250ng DNA with HindIII, 20 minutes at 37deg C and 20 min at 80deg C to inactivate. Did in new PCR machine.

DNA 250ng
enzyme 0.5
10xbuffer 2
water 9.17
Total 20


Dilute 1ul into 20ul water. run 1ul on bioanalyzer.


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