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PCR hood UV light

PCR hood UV light - PCR - Polymerase Chain Reaction Forum

PCR hood UV light - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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  #1  
Old 02-09-2011, 10:51 AM
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Default PCR hood UV light



Hi ,

since i had a lot of troubles with pcr contamination i decided to pipet my pcr reactions under a hood which is only used for PCR Pipetting.

This hood has also UV-Light and I was told to switch it on during pipetting. But does tghis make sence at all??? Waht is exactly the purpose of UV-Light under this hood???

My results were zero!!! I didnt have any PCR product at all!!! Is it possible that the UV liht is the reason??? And by teh way: UV light is harmfull for the eyes!

Would appreciate any helpful suggestions :-)
THANKS
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Old 02-09-2011, 10:59 PM
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Default Re: PCR hood UV light

You're absolutely right. You should not work under UV light. UV radiation causes the formation of thymidine dimers in the DNA rendering it useless as a template for PCR (yes UV light does not discriminate between sample and contaminant DNA and will inactivated both).

It's important to know that not only template DNA is sensitive to UV light: dNTPs are kind of resistant but the long term effects are unpredictable. Primer UV sensitivity is variable but as well unpredictable. Taq pol is very sensitive to UV damage. The only reagents that I'd recomend to irradiate with UV light are water and PCR buffers.

UV light is useful to decontaminate hood's surfaces as well pipettes, tubes, pipettes tips boxes. It's also useful to use EtOH (70%) and diluted bleach (5%) to clean the surfaces of the hood, as well as the other equipment stated before.

And by the way, UV light is not only harmful for the eyes, it is also harmful for the skin since it is an important cancer promoting agent.

Hope this helps you. The best of luck
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Old 02-10-2011, 01:39 PM
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Default Re: PCR hood UV light

Hi,

thanks very much for the broad information. Indeed i now pipeted without the uv light switched on ( just UV circulation, but dontīt know what it really means) and it worked perfectly :-)!
What about the Magnesium?
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Old 02-11-2011, 06:48 PM
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Default Re: PCR hood UV light

Oh yeah, I'm sorry, I was also refering to MgCl when I said PCR buffers. So, yes Magnessium can be irradiated, too.

Congrats on your success!
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