Team:Manchester/Safety
Safety is absolutely paramount in the scientific research area. In conducting scientific experiments, researchers should always comply to the safety measurements outlined in the Health and Safety at Work Act 1974, c.37 [1], not only to protect themselves, but also the members of the public and environment from any potential harm.
Scroll to check out the safety measurements we took!
Ethidium Bromide (EtBr)
- Hazard statements: H302, H330, H341
- Risks: Harmful if swallowed and inhaled, highly mutagenic and may be carcinogenic [3].
EtBr is commonly used in laboratories to identify the band sizes of DNA samples under ultraviolet light after being loaded onto agarose gels.
Usage of EtBr has been reduced in our lab by using SYBR Safe as a substitute to minimize the risks as stated above.
SYBR Safe
- Hazard statements: H227
- Risks: Flammable, may cause mild eye and skin irritation
Similar to EtBr, SYBR Safe enables the visualization of nucleic acid bands using agarose gels. In comparison, SYBR Safe has been identified to be 4 to 5 times less mutagenic than EtBr [4]. Thus, we substituted EtBr with SYBR Safe for most of our nucleic acid staining procedures.
ABTS
(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid))
- Hazards statements: H315, H319, H335
- Risks: May cause skin, eye and respiratory irritation.
ABTS is widely used to detect the binding activity of molecules in presence of a peroxidase enzyme (eg: HRP).
The possibility of causing skin irritation when in direct contact with ABTS indicates that ABTS may not be the best candidate for our actual AlcoPatch. For future experiments, ABTS should be substituted with other safer redox indicators.
Glucose Oxidase (GOx)
- Hazards statements: H334
- Risks: May cause skin and respiratory irritation.
GOx catalyzes the oxidation of glucose to hydrogen peroxide (H2O2) and D-glucono-δ-lactone. GOx is used in our pilot experiment to justify our proof of concept before we kick start our actual project with alcohol oxidase (AOx).
Horseradish Peroxidase (HRP)
- Hazards statements: H334, H317
- Risks: May cause skin and respiratory irritation.
HRP catalyzes the redox reaction and amplify the signals of chromogenic (eg: ABTS) and chemiluminescence (eg: luminol) substrates. The reaction is fast and visible, as shown in our pilot study .
- Design a kill switch system to minimize that the risk of releasing live E.coli organisms to the environment.
- Explore other redox indicators as alternatives for ABTS.
- Conducted by the Safety and Risk Manager of Manchester Institute of Biotechnology, Dr. Tanya Aspinall.
- Outlined the general lab safety rules, waste disposal procedures and other policies that we should abide to, including COSHH and risk assessments.
- Lab we worked in was categorized as Biosafety Level 1, according to WHO Laboratory Safety Manual [5].
- Led by one of our supervisors, Marc.
- Familiarize ourselves to work in the lab.
- Performed our lab rota under Marc’s supervision.
When in the lab:
Strictly no food and drink
Always wear a lab coat
Wear protective goggles when working with harmful chemicals
Always wear rubber gloves unless working under flame
Always work at properly ventilated area
Dispose contaminated consumables following local regulations
- Followed the shipping guidelines provided by iGEM HQ.
- Submitted our BioBricks [insert link to our BioBrick list] in pSB1C3 backbone.
- Time to get ready for the Giant Jamboree in Boston! :)
References
- Health and Safety at Work Act 1974, c.37. Available at: http://www.legislation.gov.uk/ukpga/1974/37 [Accessed 13 September 2013]
- Chart, H., Smith, H. R., La Ragione, R. M. & Woodward, M. J. (2000). An investigation into the pathogenic properties of Escherichia coli strains BLR, BL21, Dh5alpha and EQ1. Journal of Applied Microbiology, 89, 1048-1058.
- Walter, J., Caroline, J. C., Caroline, J. J. & Anjanapriya, S. (2013). Toxic and mutagenic impact of ethidium bromide traces on the growth of Escherichia coli KL226. International Journal of Current Science, 8, E28-32.
- Evenson, W. E., Boden, L. M., Muzikar, K. A. & O’Leary, D. J. (2012). 1H and 13C NMR assignments for the cyanine dyes SYBR Safe and Thiazole Orange. Journal of Organic Chemistry, 77(23), 10967-10971.
- Laboratory biosafety manual. 3rd edition. Geneva: World Health Organization; 2004. (WHO/CDS/CSR/LYO/2004. 11). Available at: http://www.who.int/csr/resources/publications/biosafety/en/Biosafety7.pdf [Accessed 14 September 2016]
