Team:UQ-Australia/Notebook

From 2009.igem.org

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(Bioprecipitation Project)
(Water Purification Project)
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=='''Water Purification Project'''==
=='''Water Purification Project'''==
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'''24/07'''<br/>
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'''24/07/09  - DNA extraction and electrophoresis'''<br/>
Mini prep DNA extractions performed for MS427-pBAD, MS427-pKKJ143 cultures incubated overnight.<br/>
Mini prep DNA extractions performed for MS427-pBAD, MS427-pKKJ143 cultures incubated overnight.<br/>
--> run on ethidium bromide gel to confirm DNA production.<br/>
--> run on ethidium bromide gel to confirm DNA production.<br/>
<br/>
<br/>
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'''23/07/09''' <br/>
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'''23/07/09   - Transformations with AG43 plasmids''' <br/>
''E. coli'' incubated overnight in duplicate under the following conditions:<br/>
''E. coli'' incubated overnight in duplicate under the following conditions:<br/>
  Strain: MS427 <br/>
  Strain: MS427 <br/>

Revision as of 14:42, 16 August 2009

Water Purification Project

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.

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

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!

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