PCR Lab Design

Jeremy Carson carsonj at ozemail.com.au
Wed Aug 30 08:03:20 EST 1995


I am in the process of setting up a lab for diagnostic PCR for bacteria. We
have two labs, one which operates with slight negative pressure - about 10 air
changes an hour.
We are proposing to prepare samples in the main lab. In this lab we have a
small downward displacement laminar flow cabinet with UV lamp. We plan on
setting up the PCR reaction in this unit. Thereafter we will transfer the tubes
to the negative pressure room and run the PCR there. We have access to a
reverse flow laminar flow unit (operator protection) where we could open the
tubes and analyse the product. We will use barrier tips, master mixes, prepare
reagents in a separate lab as basic precautions.
Comments would be appreciated on (1) will the proposed set up go a long way to
reducing contamination and (2) will HEPA filters have any capacity to bind
amplified DNA. Obviously the amplified DNA molecule is less than the 0.3 micron
pore size of the filter but I was wondering if electrostatic charge may bind
the DNA to the filter matrix.
Thanks.
Jeremy Carson




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