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Team:NTU-Singapore/Notebook/23 June 2008
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NTU@iGEM
Project
Introduction
Methodology
Team
Introduction
Team background
Notebook
Dry Lab
Introduction
ODEs used
Modeling Constructs
Parameters used
Deterministic Modeling
Sensitivity Analysis
Stochastic Modeling
Wetlab
Introduction
Materials and Equipment
Protocols
Verification of Detection & Lysis System
Our New Parts
Characterization
Introduction
Standard Promoters
Characterization of pLacI-GFP
pLacI-GFP images and clips
Characterization of pLsrA-YFP
New characterization Proposal
Parameter Estimation & Correlations
Acknowledgements
Monday 23 June
Morning (9am-1230pm)
DNA extraction (using Miniprep kit) for 21 samples that Choon Kit inoculated on
Sunday
Digestion of the above 21 plasmids by EcoRI and PstI, followed by Gel electrophoresis check after 2 hours incubation.
Digestion LacI promoter insert using SpeI and EcoRI sequentially with QiAgen PCR purification kit in between.
Digestion of RBS 0034 vector using XbaI and EcoRI sequentially and QIAgen PCR purification kit.
Chin Chong
Preapre LBA agar plates of Top10 and BL21 containing LacI-GFP and allowed to incubate overnight at 37 deg cel
Designed the Lysis gene (with Promoter and RBS) and LsrA gene and requested for Quotations from companies
Inoculate top10 and BL21 cells containing LacI-GFP and grow in two seperate tubes containing 5 ml of LBA
allowed to incubate overnight at 37 deg cel
Autoclaved MgSO4, CaCl2 and M9 salts