Team:UQ-Australia/Notebook

From 2009.igem.org

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='''Notebook'''=
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|{{#calendar: title=UQ-Australia |year=2009 | month=07}}
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|{{#calendar: title=UQ-Australia |year=2009 | month=09}}
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|{{#calendar: title=UQ-Australia |year=2009 | month=10}}
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=='''Water Purification Project'''==
 
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'''02/09/09 - Running Gel for Sandard 23 '''
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===UQ-Australia is involved in two projects:===
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* [https://2009.igem.org/Team:UQ-Australia/Notebook/Project1 '''Bioaccumulation (Mercury) Project''']
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* [https://2009.igem.org/Team:UQ-Australia/Notebook/Project2 '''Bioprecipitation Project''']
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'''01/09/09  - Agarose Gel and Nanodrop'''<br/>
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Please click on the links above to check out each Lab Notebook.  
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* Miniprepped BBa_J63010 plasmids ran on 1% Agarose gel (TAE buffer) at 200V, 70mA for 1hour. Stain: Ethidium Bromide.<br/>
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* Miniprepped BBa_J63010 plasmids (tubes A to E) analysed on nanodrop. (Data printout in IGEM_Mercury noteook)<br/>
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* New space created for BBa_J63010 tubes A to E: in "Fiona" freezer, UQ-Australia IGEM Team's ''Green'' box.<br/>
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'''28/08/09  - DNA miniprep'''<br/>
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Bon apetite!
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Mini prep DNA extractions performed for BBa_J63010 plasmid selected yesterday and incubated overnight.<br/>
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Tubes stored in "Fiona" freezer, UQ-Australia IGEM Team's orange box.<br/>
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''EDIT (01/09/09): these tubes now in IGEM''
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''Green Box ("Fiona" freezer, bottom shelf).''
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<br/>
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'''27/08/09  - stuff'''<br/>
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Could those who did work on this day upload their research notes? Cheers!<br/>
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'''26/08/09  - stuff'''<br/>
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Could those who did work on this day upload their research notes? Cheers!<br/>
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<br/>
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'''25/08/09  - stuff'''<br/>
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Could those who did work on this day upload their research notes? Cheers!
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<br/>
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'''24/07/09  - DNA extraction and electrophoresis'''<br/>
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Mini prep DNA extractions performed for MS427-pBAD, MS427-pKKJ143 cultures incubated overnight.<br/>
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--> run on ethidium bromide gel to confirm DNA production.<br/>
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Click [https://2009.igem.org/Image:UQ_MercuryMiniprep_AgaroseGel_1.png HERE] for a picture of the gel.
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'''23/07/09  - Transformations with AG43 plasmids''' <br/>
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''E. coli'' incubated overnight in duplicate under the following conditions:<br/>
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Strain: MS427 <br/>
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Plasmid: pBAD (empty plasmid -AG43) <br/>
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- 2mL LB broth <br/>
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- 2µL ampicillin <br/>
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Strain: MS427 <br/>
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Plasmid: pKKJ143 (plasmid w/ AG43) <br/>
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- 2mL LB broth <br/>
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- 2µL ampicillin<br/>
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'''17/07/09  - MG1655 Stock Preparation''' <br/>
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MG1655 ''E. coli'' grown on pure LB stock. No growth was observed using LB + ampicillin medium.<br/>
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Pelleted and resuspended in 50:50 glycerol:LB medium and stored at -80 degrees C.<br/>
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=='''Bioprecipitation Project'''==
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'''02/09/09'''
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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!
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'''14/08/09'''
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<br>''Results from transformation 2''
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*pxCK-EJ = 1 colony
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*pxCK-ES = 3 colonies
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*pxCK-EL = 27*4 colonies
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*pxCK-K = 53*4 colonies
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Plates were sealed with parafilm and stored in the -4 fridge.
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'''19/08/09'''
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Miniprep of bacterial cultures. For procedure click [https://2009.igem.org/Team:UQ-Australia/Notebook/Miniprep_procedure HERE]. Plasmids were stored in -20'C freezer (FIONA).
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'''18/08/09'''
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Colonies were picked from transformed bacteria. Protocol can be found by clicking [https://2009.igem.org/Team:UQ-Australia/Notebook/Colony_pick_procedure HERE].
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'''13/08/09'''
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A second transformation was performed on the four plasmids. Procedure slightly changed, click [https://2009.igem.org/Team:UQ-Australia/Notebook/Transformation_procedure2 HERE] to see it.
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'''11/08/09'''
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No results were given from the transformation. Transformation will have to be repeated.
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'''10/08/09'''
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Transformation of our 4 plasmids took place. They were transformed into TOP10 bacteria. A colony check will be preformed tomorrow morning.
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To see protocol, click [https://2009.igem.org/Team:UQ-Australia/Notebook/Transformation_procedure HERE]
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'''7/08/09'''
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Vectors were examined and we are planning for the best cloning strategy for the standard plasmids.
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'''6/08/09'''
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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.
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LB media was poured on petri dishes and plasmid DNA was checked again using Nanodrop.
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*pXCK-EL = 10.55 ng/uL
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*pXCK-ES = 8.43 ng/uL
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*pXCK-K = 11.73 ng/uL
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*pXCK-E/J = 2.66 ng/uL
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Plasmids were stored and transformation will be done on Monday.
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'''5/08/09'''
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Plasmids have arrived!
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*[https://2009.igem.org/Team:UQ-Australia/Notebook/pXCK-E/J pXCK-E/J]
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*[https://2009.igem.org/Team:UQ-Australia/Notebook/pXCK-K pXCK-K]
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*[https://2009.igem.org/Team:UQ-Australia/Notebook/pXCK-EL pXCK-EL]
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*[https://2009.igem.org/Team:UQ-Australia/Notebook/pXCK-ES pXCK-ES]
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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.
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A Nanodrop was used to see if the DNA was coming out of the paper. No DNA was detected, thus samples were left overnight.
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''Click on the plasmid name to look at the vector''
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Latest revision as of 13:05, 21 October 2009

Notebook

July
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6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
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3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
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7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
October
MTWTFSS
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5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31

UQ-Australia is involved in two projects:

Please click on the links above to check out each Lab Notebook.

Bon apetite!