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Team:NTU-Singapore/Notebook/9 June 2008
From 2008.igem.org
Monday 9 June
- Hung,Zhen Fu:
- Prepare plasmid DNA(in liquid form) of :
- GFP producer x 2
- RBS 0032 x 2
- RBS 0034 x 2
- Prepare digestion solutions for 6 above plasmids for later gel electrophoresis.
- Prepare plasmid DNA(in liquid form) of :
- Choon Kit:
- Amplification of T7 promoter, 4 empty plasmids ([http://partsregistry.org/wiki/index.php?title=Part:pSB1A3 pSB1A3], [http://partsregistry.org/wiki/index.php?title=Part:pSB1AT3 pSB1AT3], [http://partsregistry.org/wiki/index.php?title=Part:pSB3K3 pSB3K3], [http://partsregistry.org/wiki/index.php?title=Part:pSB1AK3 pSB1AK3]) from the Registry (as gel electrophoresis was not successful last week).
- Gel electrophoresis running for GFP producer, RBS 0032 and RBS 0034.
- Chin Chong, Darius
- Prepare things to autoclave. (LB broth, pippette tips etc)
- Prepare lactose/IPTG solution and LacI-GFP containing E coli cells for LacI-GFP characterization
- Carry out trial for LacI-GFP characterization
- 0-10mM of lactose/IPTG was investigated
- A 96 well reader is used
- Cells used in wells were 200 uL
- Varying the volumue of 100mM IPTG/lactose from 0 to 25 uL, and toping up with 50 uL to 25 uL of distilled water, stock of varying range of 0-100mM of lactose/IPTG was prepared in eppendorf tubes
- The cells were induced with the respective concentraions of IPTG/lactose only beofre the measurement commences
- Readings were planned to be collected overnight for 12hrs to have a rough guage of the maximum time possible for characterization
- However, the settings of the PC connected to the multiplate reader were not programmed to be operated for overnight, and the readings collected stopped after an hour. The settings were subsequently altered the next day morning.
