Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/10/28: 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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==mm/dd/yyyy==
==10/28/2013==
Miniprepped J04450 that I started growing on Saturday. They were in the incubator for a long time and were super red. <br>
Miniprepped J04450 that I started growing on Saturday. They were in the incubator for a long time and were super red. <br>


[[image:Pretty mRFP liq cultures.JPG|270px|Pretty mRFP]]


<br><br>
'''Miniprep'''<br>
{| {{table}}
|-
|260 ||0.034
|-
|280 ||0.021
|-
|260/280|| 1.631
|-
|ng/µL ||33.893
|-
|}
<br>
'''Restriction digest'''<br>
cut 15ul of 33ng/ul, so about 0.5ug of plasmid. Cut with E/X in a 20ul rxn for 25 minutes at 37°C<br>


<br>
Gel of cut J04450 [[image:13.10.28_J04450.png‎|J04450 E/X]]
 
<br>Cut out and put the gel slice at -20°C<br><br>
Gel image
13.10.28_J04450.png‎
 
 
PCR image
13.10.28_PCR_cropped.png‎


PCR ''need to edit this to include the next lane. lane 1 is P087/P088 and lane 2 is P089/P090, both I13522'' [[image:13.10.28_PCR_cropped.png‎|PCR]]
<br><br>
Ran another PCR reaction to text which primer wasn't working for the sender backbone. Used the two sender backbone primers, P089 and P090 with the matching F or R primer from the pLux vectors primers which were shown to work. Rxn 3 is positive control.<br>
1
:15ul reaction
:P087/P090
2
:15ul reaction
:P089/P088
3
:10ul
:P087/P088


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Latest revision as of 23:31, 26 September 2017

Project name Main project page
Previous entry      Next entry

10/28/2013

Miniprepped J04450 that I started growing on Saturday. They were in the incubator for a long time and were super red.

Pretty mRFP



Miniprep

260 0.034
280 0.021
260/280 1.631
ng/µL 33.893

Restriction digest
cut 15ul of 33ng/ul, so about 0.5ug of plasmid. Cut with E/X in a 20ul rxn for 25 minutes at 37°C

Gel of cut J04450 J04450 E/X
Cut out and put the gel slice at -20°C

PCR need to edit this to include the next lane. lane 1 is P087/P088 and lane 2 is P089/P090, both I13522 PCR

Ran another PCR reaction to text which primer wasn't working for the sender backbone. Used the two sender backbone primers, P089 and P090 with the matching F or R primer from the pLux vectors primers which were shown to work. Rxn 3 is positive control.
1

15ul reaction
P087/P090

2

15ul reaction
P089/P088

3

10ul
P087/P088


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