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

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2015/03/04 Entry for Haynes_Lab:Notebook/CRISPR_Editing)
 
(fix raw html notebook nav)
 
(4 intermediate revisions by one other user not shown)
Line 2: Line 2:
|-
|-
|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>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
|-
|-
| colspan="2"|
| colspan="2"|
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==03//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.


{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Cell type'''
| align="center" style="background:#f0f0f0;"|'''treatment'''
| align="center" style="background:#f0f0f0;"|'''rep'''
| align="center" style="background:#f0f0f0;"|'''260'''
| align="center" style="background:#f0f0f0;"|'''280'''
| align="center" style="background:#f0f0f0;"|'''260/280'''
| align="center" style="background:#f0f0f0;"|'''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
|}


<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.
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
|}
|}


__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.


Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/CRISPR_Editing/2015/03/04&oldid=1016451"