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> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |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]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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== | ==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> | |||
[[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> | |||
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> | |||
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 | ||||||||
10/28/2013Miniprepped J04450 that I started growing on Saturday. They were in the incubator for a long time and were super red.
Restriction digest Gel of cut J04450
PCR need to edit this to include the next lane. lane 1 is P087/P088 and lane 2 is P089/P090, both I13522
2
3
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