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12. Safety Warning: Picogreen must be treated as a potential mutagen as it binds
with nucleic acid, so must be handled with appropriate care. It is recommended
to use double gloves when handling the stock solution. Picogreen reagent should
be poured through activated charcoal before disposal. The charcoal must then be
incinerated to destroy the dye.
13. Safety Warning: Be careful when opening bottles containing heated agarose gel as
the solution can become superheated and inflict burns. The solution will initially
be cloudy when the agarose is suspended in the buffer, and then becomes clear
once the agarose has dissolved.
14. Safety Warning: Ethidium bromide is a potential carcinogen and mutagen.
Always wear gloves when handling ethidium bromide and equipment that may
have been in contact with ethidium bromide.
15. The gel can be run at a higher voltage (i.e., 100 V), however, the resolution
of the gel may be compromised and the DNA may be degraded under high
temperatures.
16. Ensure that your eyes and skin are adequately protected from sources of UV
light.
Acknowledgments
Thanks are due to Dr. Siddhartha Ghose, Prof. Nigel Slater, Dr. John
Woodgate and Dr. Peter Kumpalume for their guidance.
References
1. Legendre JY, Haensler J, Remy JS. Non-viral gene delivery systems (1996).
Médecine/Sciences 12, 1334–1341.
2. Ferreira GNM, Monteiro GA, Prazeres DMF, Cabral JMS. Downstream processing
of plasmid DNA for gene therapy and DNA vaccine applications (2000). Trends
Biotechnol. 18, 380–388.
3. Scherman D. Towards non viral gene therapy (2001). Bull. Acad. Natl. Med. 185,
1683–1697.
4. Davis HL. Plasmid DNA expression systems for the purpose of immunization
(1997). Curr. Opin. Biotechnol. 8, 635–640.
5. Levy MS, O’Kennedy RD, Ayazi-Shamlou P, Dunnill P. Biochemical engineering
approaches to the challenges of producing pure plasmid DNA (2001). Trends
Biotechnol. 18, 296–305.
6. Diogo MM, Ribeiro SC, Queiroz JA, Monteiro GA, Tordo N, Perrin P, Prazeres
DMF. Production, purification and analysis of an experimental DNA vaccine
against rabies (2001). J. Gene Med. 3, 577–584.
7. Johansen P, Raynaud C, Yang M, Colston MJ, Tascon RE, Lowrie DB. Antimycobacterial
immunity induced by a single injection of M. leprae Hsp65-encoding
plasmid DNA in biodegradable microspheres (2003). Immunol. Lett. 90, 81–85.