Team:Sumbawagen/Safety
From 2014.igem.org
Team:Sumbawagen/Team
From 2014.igem.org
Safety
- 1. Safety Rules in Laboratory
- 2. Safety Project
- 3. Safety Form
The basic rules in our laboratory are:
a. Everyone who join the project has been trained and pass the evaluation before join to the laboratory experiment. The training covered several topics, such as basic information about IGEM and synthetic biology, technique of DNA isolation and handling, cutting and joining of DNA, plasmid transformation, handling of the synthetic biology waste and basic rules of laboratory safety
b. Everyone who working in the laboratory should following the basic standard of laboratory safety, such as wearing safety cloth (laboratory coat, rubber gloves, and surgery mask) while working in the experiment according to the risk level type.
c. In order to minimize the contamination, we done the experiment in laminar air flow (clean bench), following the standard procedure.
d.Laboratory activity were done by the students under supervision of instructors.
c. In order to minimize the contamination, we done the experiment in laminar air flow (clean bench), following the standard procedure.
d.Laboratory activity were done by the students under supervision of instructors.
SECTION III: Results and Discussions
In this study we used BBa_I13522, which already available in our lab, instead of BBa_I20260 which is designated as device no. 1 by Measurement Interlab Study Committee. As shown in Figure 1, both devices have the same RBSs, genes and terminators, except promoters. Because our intention here is to prove whether our system using mobile phone camera is suitable for GFP measurement or not, we use BBa_I13522 for convenient reason.![](https://static.igem.org/mediawiki/2014/4/49/SBGI2.jpg)
![](https://static.igem.org/mediawiki/2014/a/a7/SBGI1.jpg)
ColorMeter program installed in mobile phone, detected RGB values, we then converted the values to luminance value to obtain only the brightness of color (http://www.scantips.com/lumin.html; accessed on Sep. 24th, 2014). The formula used is:
Luminance value = (R x 0.30) + (G x 0.59) + (B x 0.11)
Two measurements of RGB values of the tubes from different angles to correct possible lighting differences, and the calculation of luminance values are given in Table 1 and Figure 3 below.
![](https://static.igem.org/mediawiki/2014/5/5f/Measurment_Interlab_Study_1.jpg)
![](https://static.igem.org/mediawiki/2013/c/cf/Grafik_Econey.png)
Expression of GFP in theory should make the color of medium brighter thus luminance value will be lower compared to E. coli which did not express GFP. Tube no. 1, and 4 has higher luminance value than tube no. 3, which is as expectation. But tube no. 2 showed low luminance value too. The reason may be because the color differences are too small (luminance value range < 5), thus accuracy of the results was low. In our project using mRFP (data not shown), the difference of luminance value between expressed and non-expressed mRFP were about 50.
Finally to validate the expression of GFP, we used the only equipment available in our Lab, which can visualize specifically fluorescent protein including GFP ((http://en.wikipedia.org/wiki/Green_fluorescent_protein; accessed on Sep. 24th, 2014). Two observations were made, in liquid and solid states. As shown in Figure 4 (top), observation of the culture medium showed that tube no. 3 has bright yellow color compared to other tubes. The color should come from GFP expressed by E. coli BL21(DE3)/BBa_I13522 cultured with auto induction reagent. However, the color was not very significant. Thus, cells were pelleted by centrifugation and once again observed. As shown in Figure 4 (bottom), tube no. 3 showed much bright yellow color compared to other tubes. This result concluded that this tube really containsE. coli BL21(DE3)/BBa_I13522 expressing GFP.
![](https://static.igem.org/mediawiki/2013/c/ce/Picture4.png)
In conclusion, our measurement system using mobile phone camera could not detect GFP expression in liquid medium because of small differences observed by naked eyes. Thus, we used mRFP instead for our iGEM project.
Sponsors
![]( https://static.igem.org/mediawiki/2014/8/8b/All_sponsor.png)