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a doubt about PCR gel purification


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4 replies to this topic

#1 survi

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Posted 25 March 2009 - 07:16 AM

Hi everybody,
I want ask you a very simple question. Im going to use the Qiagen gel extraction kit to purify a PCR amplified product from an agarose gel with thidium bromide, and I want ask you: how do you avoid ethidium bromide contamination using this kit? Do you do "something" betwen cutting the gel and putting it into the tube?

Thank you

#2 pcrman

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Posted 25 March 2009 - 07:18 AM

Just wear gloves.

#3 survi

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Posted 25 March 2009 - 07:43 AM

thanks, but I mean how take ethidium bromide out from DNa, is it possible?

#4 almost a doctor

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Posted 25 March 2009 - 07:50 AM

thanks, but I mean how take ethidium bromide out from DNa, is it possible?


As far as I know the kit does that for you. Between gel and kit you dont need to do anything, and then your DNA is ready for any downstream use you need.

:)

#5 bochum

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Posted 25 March 2009 - 08:22 AM

Dont worry about Ethidium bromide it will go off during the washing step of PCR product purification. EtBr never form any strong bounding with DNA so simple washing is enough to remove EtBr from PCR...so there will no Etbr in your pcr product...
With Regards
Martin
Career In Life Science




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