Miniprep of RBS-RFP-Terminator, Assembly 1 (J23100 + GFP), pBAD reverse and Amp/Tet Construction Plasmid
- Day cultures of 5.6mL media and cells innoculated from plates were started at 10:50
- RBS-RFP-Terminator, Assembly 1 and pBAD reverse were all grown using LB-Amp
- Amp/Tet Construction Plasmid was grown on LB-Tet
- Three cultures of Assembly 1 were started because the colonies on the plates did not have a noticible green flouresence
- Miniprepped using a Invitrogen ChargeSwitch Pro Filter Plasmid Miniprep kit after 7 hours culture
- Did not observe many cells after being spun down in a Galaxy 16 centrifuge
- Used 50uL of Elution Buffer instead of 100uL as wanted as high of a concentration as possible as the number of cells is less then usual (was not inncubated as long as usual)
- Eric started overnight cultures of all 4 plasmids in case we need to do another miniprep tommorow
==Preparation of Miniprep ON Cultures)
- Innoculated the following cultures of E. coli (done with Charles)
Plasmid/Part
|
Strain of E. coli
|
Antibiotic
|
Description
|
pSB1At3 |
DB3.1 |
Tetracycline |
Tet construction plasmid
|
|
DB3.1 |
Ampicillin |
P_BAD Reverse
|
|
DB3.1 |
Ampicillin |
RBS-RFP-Terminator
|
|
DB3.1 |
Ampicillin |
Assembly 1
|
- Took a colony and innoculated fresh 6 mL LB-antibiotic
- Incubated at AMBL 37ºC incubator
- Tomorrow: miniprep the cultures and make glycerol stocks of these.
Ampicillin - New stocks testing
- Made a new stock of Amp-100
- 20 mL sH2O + 2.0755 g ampicillin sodium salt
- Filter sterilized
- Stored in 2x 15 mL culture tubes (1000X stock)
- Tested by spread plating 20µL of stock
|