Revision as of 18:48, 22 August 2018

OriginALS

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InteLab

Our team chose to participate in the Fifth International InterLab Measurement Study in synthetic biology.
The goal of the iGEM InterLab Study is to identify and correct the sources of systematic variability in synthetic biology measurements, so that eventually, measurements that are taken in different labs will be no more variable than measurements taken within the same lab.
In this study we want to reduce lab-to-lab variability in fluorescence measurements by normalizing it to absolute cell count or colony-forming units (CFUs) instead of OD.
The fluorescence in the bacteria in our experiments occurs due to the presence of a gene that encodes a green fluorescent protein: GFP.
In cell and molecular biology, the GFP gene is frequently used as an expression reporter protein. Reporter genes are often used as an indication that a certain gene has been internalized or expressed in cells or organisms.

Equipment

All of our measurements were taken in a plate reader that can read both absorbance and fluorescence (Neoteck-Tecan-INFINITE M100) without a pathlength correction. The temperature was set to 25̊C, under shaking with a duration of 3 sec and an amplitude of 2 mm. We went through a tutorial and learned how to operate the machine.
In addition, we used the following reagents (Partially supplied by iGEM):

1.0 ml LUDOX CL-X
150 μL Silica Bead (microsphere suspension)
Fluorescein (powder, in amber tube)
iGEM Parts Distribution Kit Plates
1 x PBS (phosphate buffered saline, pH 7.4 - 7.6)
ddH2O
Competent cells (Escherichia coli DH5α)
LB (Luria Bertani) media
Chloramphenicol
96 well plates, black with clear flat bottom

Safety procedures

As part of an ongoing work in a biological laboratory, safety gear was used to work with bacteria. The work was done with closed shoes, long pants, gloves and lab coats.
In order to maintain a clean environment, and sterility between the samples the work was performed next to a flame, and all the disposable parts were thrown after a single use into a biological waste bin.
Before and after work, work surfaces were sterilized with 70% alcohol.

Protocols

Calibration 1: OD600 Reference point - LUDOX Protocol

100 μL LUDOX were added into wells A1, B1, C1, D1.
100 μLof ddH2O were added into wells A2, B2, C2, D2

Company	Contact	Country
Alfreds Futterkiste	Maria Anders	Germany
Centro comercial Moctezuma	Francisco Chang	Mexico
Ernst Handel	Roland Mendel	Austria
Island Trading	Helen Bennett	UK
Laughing Bacchus Winecellars	Yoshi Tannamuri	Canada
Magazzini Alimentari Riuniti	Giovanni Rovelli	Italy

host a group of high-school

Our project would not have succeeded without our Human Practices. Our goals and implementation were all a result of involvement in the communities around us and sharing our ideas through discussions with experts. Science cannot proceed unaided and cannot succeed without public support. As a final gesture, we were delighted to host a group of high-school students in our lab. We had a great time explaining to them about iGEM, our project, and our lab. Although we ourselves as undergraduate students are only at the beginning of our scientific careers, it was wonderful to transfer the knowledge we have gathered to a new generation.