Team:UQ-Australia/Notebook

Water Purification Project

17/07/09 - MG1655 Stock Preparation
MG1655 E. coli grown on pure LB stock. No growth was observed using LB + ampicillin medium.
Pelleted and resuspended in 50:50 glycerol:LB medium and stored at -80 degrees C.

23/07/09
E. coli incubated overnight under the following conditions:

Strain: MS427
Plasmid: pBAD (empty plasmid -AG43)
- 2mL LB broth
- 2µL ampicillin

Strain: MS427
Plasmid: pKKJ143 (plasmid w/ AG43)
- 2mL LB broth
- 2µL ampicillin

24/07
Mini prep DNA extractions performed for MS427-pBAD, MS427-pKKJ143 cultures incubated overnight.
--> run on ethidium bromide gel to confirm DNA production.

Bioprecipitation Project

5/08/09

Plasmids have arrived!

• pXCK-E/J
• pXCK-K
• pXCK-EL
• pXCK-ES

We have put them into 150 mL of TE buffer and left overnight for the paper to dissolve and later extract the plasmid from the solution.

A Nanodrop was used to see if the DNA was coming out of the paper. No DNA was detected, thus samples were left overnight.

Click on the plasmid name to look at the vector

6/08/09

Plasmid solutions were left overnight and in the morning checked again with Nanodrop. No DNA was measured. Samples were put in the waterbath at 37 degrees and left for a couple of hours. If Nanodrop does not work after the waterbath we will still do transformation since we only need few plasmids for the transformation to work. Nanodrop may not have been able to quantify such a small concentration of plasmids.

LB media was poured on petri dishes and plasmid DNA was checked again using Nanodrop.

• pXCK-EL = 10.55 ng/uL
• pXCK-ES = 8.43 ng/uL
• pXCK-K = 11.73 ng/uL
• pXCK-E/J = 2.66 ng/uL

Plasmids were stored and transformation will be done on Monday.

7/08/09 Vectors were examined and we are planning for the best cloning strategy for the standard plasmids.

10/08/09 Transformation of our 4 plasmids took place. They were transformed into TOP10 bacteria. A colony check will be preformed tomorrow morning.

To see protocol, click HERE

11/08/09 No results were given from the transformation. Transformation will have to be repeated.