Template:SBB-Protocols Assay1: Difference between revisions
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* Replate 16 constructs on new plates at the same time. | * Replate 16 constructs on new plates at the same time. | ||
* Plate DH10B, pBca9145-Bca9494, and pBca9495CA-Bca1144 at the same time. | * Plate DH10B, pBca9145-Bca9494<font color=red> (That's OVER 9000!!) </font>, and pBca9495CA-Bca1144 at the same time. | ||
* Grow plates overnight. | * Grow plates overnight. | ||
Revision as of 12:34, 15 April 2009
Growth Assay Protocol
- Replate 16 constructs on new plates at the same time.
- Plate DH10B, pBca9145-Bca9494 (That's OVER 9000!!) , and pBca9495CA-Bca1144 at the same time.
- Grow plates overnight.
- Pick 5 colonies from each sample.
- Grow to saturation in 96 well blocks with 400 uL LB media in each well.
- Then from each of the 5 unique liquid cultures, make an arabinose sample and a non-arabinose sample.
- Add 50 uL of LB media per well in 384 well plate.
- Add 1 uL of cell sample to each well.
- Add 1000x Arabinose to 5 out of the 10 wells for each construct.
- Place plate in Tecan and run OD measurements every 10 minutes.
Schedule
- For the first run-through, we'll do the 3 controls.
- Get one of the
amazingGSI's to pick 5 colonies of each control Sunday night, suspending each colony in 400 uL LB media in 96 well plate.
Monday
- Run the assay, starting from the 384 well plate step.
- Plate all the constructs and the controls.
Tuesday
- Pick 5 colonies from each plate and grow on the 96 well block to saturation.
Wednesday
- Run the assay from the 384 well plate step.
