Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/04/15: 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;">Transformation Results for DBN001_pSB1A3</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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==Transformation Results for mCherry vector + gBlock==
==Overview==
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Here are the colonies observed after one weekend grown at room temperature. I'll also be doing a plasmid miniprep of mCh_pSB1A3 so that I have more stocks of the cloning vector.


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I'm going to set up two cultures for the two colonies on the sample plate, but am not very confident of the success of the ligation given the colonies on the negative control.
I'm going to set up two cultures for the two colonies on the sample plate, but am not very confident of the success of the ligation given the colonies on the negative control.


==Plasmid Miniprep of mCherry Vector==
==mCh_pSB1A3 Plasmid Miniprep==


<b>--DNA Concentration--</b>
<b>--DNA Concentration--</b>

Revision as of 11:18, 22 April 2015

Transformation Results for DBN001_pSB1A3 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Overview

Here are the colonies observed after one weekend grown at room temperature. I'll also be doing a plasmid miniprep of mCh_pSB1A3 so that I have more stocks of the cloning vector.

Plate # colonies
positive control many dozens
sample 2
negative control 2

I'm going to set up two cultures for the two colonies on the sample plate, but am not very confident of the success of the ligation given the colonies on the negative control.

mCh_pSB1A3 Plasmid Miniprep

--DNA Concentration--

Sample ID Concentration (ng/µL) St Dev (ng/µL)
mCh Vector 1 392.1 15.5
mCh Vector 2 315.5 18.1
mCh Vector 3 319.7 6.3
mCh Vector 4 375.1 7.1

--Restriction Digest--

Reagent Volume
DNA 600ng (1.9 - 1.5 µL)
10x FastDigest Buffer 3
EcoRI 1
PstI 1
Water 23
Total 30

Digest for 15 minutes at 37°C.


--Gel Electrophoresis--

Expected size fragments from EcoRI/PstI digest: 2114, 980



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