Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/10/28: Difference between revisions
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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> | Miniprepped J04450 that I started growing on Saturday. They were in the incubator for a long time and were super red. <br> | ||
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<br><br> | <br><br> | ||
'''Miniprep'''<br> | '''Miniprep'''<br> | ||
{| {{table}} | |||
|- | |||
|260 ||0.034 | |||
|- | |||
|280 ||0.021 | |||
|- | |||
|260/280|| 1.631 | |||
|- | |||
|ng/µL ||33.893 | |||
|- | |||
|} | |||
<br> | |||
'''Restriction digest'''<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> | 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> | ||
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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]] | 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> | <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. | 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 | 1 | ||
:15ul reaction | :15ul reaction |
Revision as of 16:17, 2 December 2013
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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
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