Team:UQ-Australia/Notebook

Water Purification Project

01/09/09 - Agarose Gel and Nanodrop

• Miniprepped BBa_J63010 plasmids ran on 1% Agarose gel (TAE buffer) at 200V, 70mA for 1hour. Stain: Ethidium Bromide.
  
• Miniprepped BBa_J63010 plasmids (tubes A to E) analysed on nanodrop. (Data printout in IGEM_Mercury noteook)
  
• New space created for BBa_J63010 tubes A to E: in "Fiona" freezer, UQ-Australia IGEM Team's Green box.
  

28/08/09 - DNA miniprep
Mini prep DNA extractions performed for BBa_J63010 plasmid selected yesterday and incubated overnight.
Tubes stored in "Fiona" freezer, UQ-Australia IGEM Team's orange box.

EDIT (01/09/09): these tubes now in IGEM
Green Box ("Fiona" freezer, bottom shelf).

27/08/09 - stuff
Could those who did work on this day upload their research notes? Cheers!

26/08/09 - stuff
Could those who did work on this day upload their research notes? Cheers!

25/08/09 - stuff
Could those who did work on this day upload their research notes? Cheers!

24/07/09 - DNA extraction and electrophoresis
Mini prep DNA extractions performed for MS427-pBAD, MS427-pKKJ143 cultures incubated overnight.
--> run on ethidium bromide gel to confirm DNA production.
Click HERE for a picture of the gel.

23/07/09 - Transformations with AG43 plasmids
E. coli incubated overnight in duplicate 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

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.

Bioprecipitation Project

02/09/09 For the last couple of weeks we have been trying to design primers in order to put them into the iGEM standard plasmids. Since restriction sites are against us, we are using mutagenesis in order to bypass this problem. Primers were ordered TODAY!

14/08/09
Results from transformation 2

• pxCK-EJ = 1 colony
• pxCK-ES = 3 colonies
• pxCK-EL = 27*4 colonies
• pxCK-K = 53*4 colonies

Plates were sealed with parafilm and stored in the -4 fridge.

18/08/09 Colonies were picked from transformed bacteria. Protocol can be found by clicking HERE.

13/08/09 A second transformation was performed on the four plasmids. Procedure slightly changed, click HERE to see it.

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

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

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

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.

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