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 Images and Videos from the Characterization Lab 1.1 pLacI-GFP part:BBa_J04430 found to express RFP instead 2 Image results obtained from fluorescence microscope 2.1 Weak promoter [http://partsregistry.org/Part:BBa_J23150 BBa_J23150] 2.2 Medium promoter [http://partsregistry.org/Part:BBa_J23151 BBa_J23151] 2.3 Strong promoter [http://partsregistry.org/Part:BBa_J23102 BBa_J23102] 2.4 Standard promoter with GFP reporter [http://partsregistry.org/Part:BBa_I20260 BBa_I20260] 3 Video obtained from fluorescence microscope

Images and Videos from the Characterization Lab

pLacI-GFP part:BBa_J04430 found to express RFP instead

The below pictures were taken after the 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. In order to solve this problem the pLac promoter (R0010) was ligated with the GFP (E0040), allowing us to proceed further with the characterization experiments.

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 of the various standards (Low, Medium, High & Standard) that iGEM Headquarters has provided for us in the [http://partsregistry.org/Measurement/SPU/Learn iGEM Newsletter].

Weak promoter [http://partsregistry.org/Part:BBa_J23150 BBa_J23150]

BBa_J23150 Before Excitation

BBa_J23150 After Excitation

Medium promoter [http://partsregistry.org/Part:BBa_J23151 BBa_J23151]

BBa_J23151 Before Excitation

BBa_J23151 After Excitation

Strong promoter [http://partsregistry.org/Part:BBa_J23102 BBa_J23102]

BBa_J23102 Before Excitation

BBa_J23102 After Excitation

Standard promoter with GFP reporter [http://partsregistry.org/Part:BBa_I20260 BBa_I20260]

BBa_I20260Before Excita

BBa_I20260After Excitation

The above pictures showed the green fluorescence protein was produced by Top 10 competent cells that were taken under blue light excitation. These cells contained the different standard promoters with GFP and they have been allowed to grow to an OD of 1.2 before a snap shot was taken with the microscope.

Video obtained from fluorescence microscope

Video of E coli under normal white light

Video of E coli under blue light excitation