Manipulation with PCR components - Laminar, PCR box? (Jul/19/2005 )
What is the best place for preparation of PCR mix: laminar, PCR-box, just table or something else?
My experience has been PCR contamination is generally not too much of a problem until you have cloned your PCR fragment and done a miniprep. After a miniprep everything in the lab seems to become contaminated.
I think a PCR box will be good. I think is more important that you have 3 physicalil seperated placed for your pre-PCR, PCR and post PCR.
We have a seperate room with laminar flow for prePCR work, a room for adding template to the mixes, a room where we have our PCR-machines and another flow for adding PCR to new mixes (in case nested PCR is performed, after adding mixes UV light is put on for at least 15 minutes in the cabin) and a post PCR room with cyclers for sequencing/inner PCR. For the last two mentioned, the mixes are also prepared in the prePCR laminar flows.