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

Contents 1 pLacI-GFP part:BBa_J04430 found to express RFP instead 2 Image Results obtained from fluorescence microscope 2.1 Standard 102 2.2 Standard 150 2.3 Standard 151 2.4 Standard 260 3 Video obtained from fluorescence microscope

pLacI-GFP part:BBa_J04430 found to express RFP instead

The below pictures were taken after the biobrick part: BBa_J04430 was transformed into Top 10 competent cells. They clearly showed, either under normal light or UV light, the cells appears red or glow red respectively. This is contrary to what was expected, as we would expect it to glow green under UV light. This problem was highlighted to the iGEM Headquarters. We decided to ligate the pLac promoter (R0010) with the GFP (E0040) ourselves and the result was good this time round as the cells glow green under UV light as expected.

Cells appears pinkish even under normal light

Cells glow PINK under UV light

Image Results obtained from fluorescence microscope

Below are Images obtained from the microscope on the various standards that iGEM MIT has provided for us this year.

Standard 102

102 Normal

102 GFP

Standard 150

150 Normal

150 GFP

Standard 151

151 Normal

151 GFP

Standard 260

260 Normal

260 GFP

The above picture shows the green fluorescence produced by Top 10 ecoli cells under blue light excitation. These Ecoli cells contain the standard promoter 260 with GFP and thye have been allowed to grow to an OD of 1.2 before a snap shot is taken.

Video obtained from fluorescence microscope

Video of E coli under normal white light

Video of E coli under blue light excitation