Protocol Online logo
Top : Forum Archives: : Real-Time PCR

UV steriliser - (Oct/03/2006 )

hi guys,

does anybody know of a supplier/brand/model for a UV steriliser i could use to make sure my pipettes aren't contaminated after every real-time i run??? i can only seem to find the ones used in hairdressing salons when i web search!!!

regards

-flashboy-

May use filter tips for real-time PCR to avoid contamination.

-Minnie Mouse-

hi minnie,

i currently use PCR grade filter tips, as well as nuclease free water, and perform my PCR in a sterile hood which gets swabbed down before and after with sodium hydroxide... sadly, the pipettes we all use for PCR will more than likely be smothered in DNA from each run performed so ideally they should be sterilised [i've heard that using glycine-HCL buffer pH2.0 or UV irradiation will work]... i justneed to know where to get the UV irradiator!

thanks anyway

-flashboy-

We were thinking to buy a solution from Ambion called DNAzap that is supposed to remove DNA

-dnafactory-

yeah, DNAzap or DNa eraser solutions will remove contamination from DNA (just as RNAZap and Rnaseaway will do the same for RNA) but these solutions are better suited to decontaminating surfaces... my advice is to save money and clean your surfaces with 20mM NaOH which does the same thing....

this is kind of possible to use with pipettes but its better to UV irradiate as it involves less contact with solutions. VWR used to provide a UV irradiator but don't anymore, i might have to buy the kind of thing you see in a hair salon!!!

-flashboy-

Seems like you have thought of most things, but I thought I would suggest one more:

You might contact the manufacturer of your sterile hood and see if you can get a UV lamp that fits inside. You can leave the UV lamp on when not in use, and also sterilize your pipets by putting them up-side-down in a beaker.

In any case, good luck finding what you need!

-soluene-

UV is overrated. It has to actually hit the surface to do any good. Your pipettors won't be treated effectively by UV in any case. Use filter tips, gloves, and keep your lab coat away from the sample setup. I'd suggest that if you have continuing problems that the difficulty is not in the pipettors but rather somewhere else. Don't autoclave tubes and tips -- use them straight from the box. Test each component of the setup individually with a working negative control by adding small amounts of each tested component to the negative control.

-phage434-