Team:UQ-Australia/Notebook
From 2009.igem.org
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=='''Mercury Vacuum Project'''== | =='''Mercury Vacuum Project'''== | ||
+ | '''15/09/09 - Repeat Transformation of BBaI716101'''<br/> | ||
+ | The overnight transformation from 14/09/09 was unsuccessful and no growth was observed on plates. It is probable that the cell death plasmid component (Part BBa_P1010) is functioning normally as it was believed that this component was 'faulty'. The same protocol as on 14/09/09 was used for the transformation with an extra positive control for comparison. A well characterised plasmid PUC18 (ampicillin resistance) was used as the positive control. | ||
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'''14/09/09 - Transformation of Plate 1, Well C'''<br/> | '''14/09/09 - Transformation of Plate 1, Well C'''<br/> | ||
PLasmid BBa_I716101 with Part BBa_P1010 was transformed into XL-10 Gold Ultracompetent cells via heat shock and left in ampicillin plates at 37C overnight. DNA from plate 1, well C was stored in the Green iGEM box ('Fiona').<br/> | PLasmid BBa_I716101 with Part BBa_P1010 was transformed into XL-10 Gold Ultracompetent cells via heat shock and left in ampicillin plates at 37C overnight. DNA from plate 1, well C was stored in the Green iGEM box ('Fiona').<br/> |
Revision as of 05:01, 17 September 2009
Mercury Vacuum Project15/09/09 - Repeat Transformation of BBaI716101 14/09/09 - Transformation of Plate 1, Well C 02/09/09 - Running Gel for Standard 23 01/09/09 - Agarose Gel and Nanodrop
EDIT (01/09/09): these tubes now in IGEM Green Box ("Fiona" freezer, bottom shelf).
27/08/09 - Colony Picking 26/08/09 - Transformation of Plasmid BBa_J63010 with part BBa_J04450 --->Digest Gel of BBa_I716101 with pGfa2cLac2 control 25/08/09 - In-gel Extraction and Overnight Digests 24/07/09 - DNA extraction and electrophoresis 23/07/09 - Transformations with AG43 plasmids Strain: MS427
Strain: MS427 20/08/09 - Plasmid Digestion 19/08/09 - Miniprepping and Running PCR Gel + Nanodrop 18/08/09 - PCR Preparation and Picking Cultures 24/07/09 - Extraction of DNA from E.coli M5427
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Bioprecipitation Project08/09/09 Our Primers have arrived today, this means we can start to put our four genes into the standard plasmids! 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! 19/08/09 Miniprep of bacterial cultures. For procedure click HERE. Plasmids were stored in -20'C freezer (FIONA). 18/08/09 Colonies were picked from transformed bacteria. Protocol can be found by clicking HERE. 14/08/09
Plates were sealed with parafilm and stored in the -4 fridge. 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.
To see protocol, click HERE
LB media was poured on petri dishes and plasmid DNA was checked again using Nanodrop.
Plasmids were stored and transformation will be done on Monday.
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 |