Difference between revisions of "Team:USP-EEL-Brazil/InterLab"

Revision as of 02:06, 14 October 2018

IGEM is an international synthetic biology competition, which aims to set up standard protocols that work in any laboratory on the planet. To manage this task, Interlab Study was created, inviting all team to participate to complete this goal. Until we reach this point, synthetic biology will not be able to achieve its full potential as an engineering discipline, because labs will not be able to reliably build upon others’ work.

This year, Interlab study became a requisite to reach bronze medal. This sense, every team received a kit with the same materials, with which they were oriented to follow the same protocols. Through the results obtained by each team, the samples will be compared, and then, the community could get trustable info. Analyzing the info, some adjustments will be made in the protocols or even new protocols can be created to improve the synthetic biology standardization.

The Interlab consist in making measures of the Green Fluorescent Protein (GFP) and use the values to determine the feasible cell population. In order to determine cell population, scientists usually use spectrophotometry to measure cells culture absorbance in 600 nm, and through it, the optical density, OD, can be obtained. So, theoretically, the higher cells number the higher OD. However, the values of OD obtained are under high variability, because of different calibration and precision between labs equipment, different glassware size, and others lab-to-lab factors. In addition, through this methodology, it isn’t possible to separate dead feasible cells.

In this sense, to respond to all the problems above, the methodology of measuring GFP is another way to determine feasible cells, once the dead ones will not express GFP. Through the GFP measurement methodology, the Measurement Committee wants our help to answer the following question:

‘’Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD? ‘’

Materials and Methods

In order to make the experiments, we followed the method in the link below: (colocar o link)

The plasmids used are available in the iGEM kit:

Device Part Number Plate Location

Negative control BBa_R0040 Kit Plate 7 Well 2D

Positive control BBa_I20270 Kit Plate 7 Well 2B

Test Device 1 BBa_J364000 Kit Plate 7 Well 2F

Test Device 2 BBa_J364001 Kit Plate 7 Well 2H

Test Device 3 BBa_J364002 Kit Plate 7 Well 2J

Test Device 4 BBa_J364007 Kit Plate 7 Well 2L

Test Device 5 BBa_J364008 Kit Plate 7 Well 2N

Test Device 6 BBa_J364009 Kit Plate 7 Well 2P

Equipment used for measurements: TECAN INFINITY M200 PRO Plate used for measurements: Corning^® Costar Clear Polystyrene 96-Well Plates

Results

Interlab

https://2017.igem.org/Team:ColumbiaNYC/InterLab

https://2017.igem.org/Team:NUS_Singapore/InterLab

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 <p>IGEM is an international synthetic biology competition, which aims to set up standard protocols that work in any laboratory on the planet. To manage this task, Interlab Study was created, inviting all team to participate to complete this goal. Until we reach this point, synthetic biology will not be able to achieve its full potential as an engineering discipline, because labs will not be able to reliably build upon others’ work. </p>
-<p>This year, Interlab study became a requisite to reach bronze medal. This sense, every team received a kit with the same materials, with which they were oriented to follow the same protocols. Through the results obtained by each team, the samples will be compared, and then, the community could get trustable infos. Analyzing the infos, some adjust will be made in the protocols or even new protocols can be created to improve the synthetic biology standardization.
+<p>This year, Interlab study became a requisite to reach bronze medal. This sense, every team received a kit with the same materials, with which they were oriented to follow the same protocols. Through the results obtained by each team, the samples will be compared, and then, the community could get trustable info. Analyzing the info, some adjustments will be made in the protocols or even new protocols can be created to improve the synthetic biology standardization.
 </p>
-<p>The Interlab consist in making measures of the Green Fluorescent Protein (GFP) and use the values to determine the feasible cell population. In order to determine cell  population, scientists usually use spectrophotometry to measure cells culture absorbance in 600 nm, and through it the optical density, OD, can be obtained. So, theoretically, the higher cells number the higher OD. However, the values of OD obtained are under high variability, because of differents calibration and precision between labs equipments, differents glassware size and others lab-to-lab factors. In addition, through this methodology, it isn’t possible to separate dead feasible cells.
+<p>The Interlab consist in making measures of the Green Fluorescent Protein (GFP) and use the values to determine the feasible cell population. In order to determine cell population, scientists usually use spectrophotometry to measure cells culture absorbance in 600 nm, and through it, the optical density, OD, can be obtained. So, theoretically, the higher cells number the higher OD. However, the values of OD obtained are under high variability, because of different calibration and precision between labs equipment, different glassware size, and others lab-to-lab factors. In addition, through this methodology, it isn’t possible to separate dead feasible cells.
 </p>
-<p>In this sense, to respond all the problems above, the methodology of measuring GFP is another way to determine feasible cells, once the dead ones will not express GFP. Through the GFP measurement methodology, the Measurement  Committee wants our help to answer the following question:
+<p>In this sense, to respond to all the problems above, the methodology of measuring GFP is another way to determine feasible cells, once the dead ones will not express GFP. Through the GFP measurement methodology, the Measurement  Committee wants our help to answer the following question:
 </p>
 <p>‘’Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD? ‘’