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>}}
|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}}}}]]}}
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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==
 
Decent yield for all four colonies picked.


<b>--DNA Concentration--</b>
<b>--DNA Concentration--</b>
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[[Image:2015-04-15_mCherry_vector_digest_annotated.png]]
[[Image:2015-04-15_mCherry_vector_digest_annotated.png]]


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




Latest revision as of 00:54, 27 September 2017

Transformation Results for DBN001_pSB1A3 Main project page
Previous entry      Next entry

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

Decent yield for all four colonies picked.

--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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