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

From OpenWetWare
Jump to navigationJump to search
(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. ##### -->
==02//2015==
==03/03/2015==
Checked concentrations on minipreps from last year of Lonza pMax GFP from Krishna. very low so JW transformed the original along with pMax GFP 6.2 and KAH60.
Checked concentrations on minipreps from last year of Lonza pMax GFP from Krishna. very low so JW transformed the original along with pMax GFP 6.2 and KAH60.
{| {{table}}
{| {{table}}
Line 21: Line 21:
| pMax GFP||0.031||0.019||1.639||30.851
| pMax GFP||0.031||0.019||1.639||30.851
|}
|}
 
<br><br>
 
Ran PCR products from P151/158 on gel. Only single product for each band.
<br><br>
Sequencing:<br>Need to purify. Will purify 25ul and elute into 35 ul.<br>
Purification results:
{| {{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||Untreated||1||0.035||0.017||2.006||35.007
|-
| Luc14||g046||1||0.032||0.016||2.02||31.691
|-
| Luc14||Untreated||2||0.045||0.022||2.034||44.797
|-
| Luc14||Untreated||3||0.046||0.024||1.938||46.277
|-
| Gal4-EED +dox||Untreated||1||0.04||0.019||2.099||40.212
|-
| Gal4-EED +dox||Untreated||2||0.043||0.023||1.885||42.773
|-
| Gal4-EED +dox||Untreated||3||0.044||0.023||1.93||43.675
|-
| Luc14||g046||2||0.03||0.015||2.086||30.259
|-
| Luc14||g046||3||0.031||0.015||2.113||31.368
|-
| Gal4-EED +dox||g046||1||0.035||0.017||2.042||35.122
|-
| Gal4-EED +dox||g046||2||0.039||0.019||2.05||38.501
|-
| Gal4-EED +dox||g046||3||0.041||0.022||1.897||41.336
|-
| Luc14||Untreated||1||0.032||0.016||1.949||31.937
|-
| Luc14||g046||1||0.03||0.015||1.94||29.645
|}
<br>
Sent for sequencing at biodesign with P176
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Label'''
| align="center" style="background:#f0f0f0;"|'''Cell type'''
| align="center" style="background:#f0f0f0;"|'''treatment'''
| align="center" style="background:#f0f0f0;"|'''rep'''
| align="center" style="background:#f0f0f0;"|'''PCR primer'''
|-
| 1||Luc14||Untreated||1||P176
|-
| 2||Luc14||Untreated||2||P176
|-
| 3||Luc14||Untreated||3||P176
|-
| 4||Gal4-EED+dox||Untreated||1||P176
|-
| 5||Gal4-EED+dox||Untreated||2||P176
|-
| 6||Gal4-EED+dox||Untreated||3||P176
|-
| 7||Luc14||g034||1||P176
|-
| 8||Luc14||g034||2||P176
|-
| 9||Luc14||g034||3||P176
|-
| 10||Gal4-EED+dox||g034||1||P176
|-
| 11||Gal4-EED+dox||g034||2||P176
|-
| 12||Gal4-EED+dox||g034||3||P176
|-
| 13||Luc14||g046||1||P176
|-
| 14||Luc14||g046||2||P176
|-
| 15||Luc14||g046||3||P176
|-
| 16||Gal4-EED+dox||g046||1||P176
|-
| 17||Gal4-EED+dox||g046||2||P176
|-
| 18||Gal4-EED+dox||g046||3||P176
|}
<br><br>
Ran PCR with P151/158 on g023, g045, g048 PCR with P197/198 diluted 1:1000 from yesterday. Ran on gel.
<!-- ##### 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

03/03/2015

Checked concentrations on minipreps from last year of Lonza pMax GFP from Krishna. very low so JW transformed the original along with pMax GFP 6.2 and KAH60.

Sample Read# 260 280 260/280 ng/µL
pMax GFP 0.026 0.015 1.736 26.294
pMax GFP 0.023 0.013 1.766 23.175
pMax GFP 0.031 0.019 1.639 30.851



Ran PCR products from P151/158 on gel. Only single product for each band.

Sequencing:
Need to purify. Will purify 25ul and elute into 35 ul.
Purification results:

Cell type treatment rep 260 280 260/280 ng/µL
Luc14 Untreated 1 0.035 0.017 2.006 35.007
Luc14 g046 1 0.032 0.016 2.02 31.691
Luc14 Untreated 2 0.045 0.022 2.034 44.797
Luc14 Untreated 3 0.046 0.024 1.938 46.277
Gal4-EED +dox Untreated 1 0.04 0.019 2.099 40.212
Gal4-EED +dox Untreated 2 0.043 0.023 1.885 42.773
Gal4-EED +dox Untreated 3 0.044 0.023 1.93 43.675
Luc14 g046 2 0.03 0.015 2.086 30.259
Luc14 g046 3 0.031 0.015 2.113 31.368
Gal4-EED +dox g046 1 0.035 0.017 2.042 35.122
Gal4-EED +dox g046 2 0.039 0.019 2.05 38.501
Gal4-EED +dox g046 3 0.041 0.022 1.897 41.336
Luc14 Untreated 1 0.032 0.016 1.949 31.937
Luc14 g046 1 0.03 0.015 1.94 29.645


Sent for sequencing at biodesign with P176

Label Cell type treatment rep PCR primer
1 Luc14 Untreated 1 P176
2 Luc14 Untreated 2 P176
3 Luc14 Untreated 3 P176
4 Gal4-EED+dox Untreated 1 P176
5 Gal4-EED+dox Untreated 2 P176
6 Gal4-EED+dox Untreated 3 P176
7 Luc14 g034 1 P176
8 Luc14 g034 2 P176
9 Luc14 g034 3 P176
10 Gal4-EED+dox g034 1 P176
11 Gal4-EED+dox g034 2 P176
12 Gal4-EED+dox g034 3 P176
13 Luc14 g046 1 P176
14 Luc14 g046 2 P176
15 Luc14 g046 3 P176
16 Gal4-EED+dox g046 1 P176
17 Gal4-EED+dox g046 2 P176
18 Gal4-EED+dox g046 3 P176



Ran PCR with P151/158 on g023, g045, g048 PCR with P197/198 diluted 1:1000 from yesterday. Ran on gel.


Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/CRISPR_Editing/2015/03/03&oldid=1016441"