Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/10/28

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mm/dd/yyyy

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

Pretty mRFP



Miniprep
33ng/ul.
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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