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Polymerase Chain Reaction technique question?

Polymerase Chain Reaction technique question? - PCR - Polymerase Chain Reaction Forum

Polymerase Chain Reaction technique question? - 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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Old 09-06-2011, 02:16 AM
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Default Polymerase Chain Reaction technique question?



The question is will I be able to know if my strategy to separate the DNA from other cellular components (proteins, starch, etc) has succeeded for failed?

Basically, I extract the DNA from the cell using buffer and enzyme. My strategy to separte the DNA is that I add ethanol in high salt condition so that the DNA will bind to the silica gel in chromatography column. Then I wash the sample to remove the cellular stuff, centrifuge, and dilute the DNA with water. Then I do the first round of PCR.

so will I know if that strategy has worked? I'm thinking yes because if the separation failed then nothing will be amplified in PCR and we would know that right?
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Old 09-06-2011, 08:52 PM
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Default Re: Polymerase Chain Reaction technique question?

Yes you can say with PCR that you isolated DNA. But you not able to say whether Your DNA purified from Protein other competent. Some PCR work even if you one copy of DNA in a sample which has grams of Protein. some time People use to do PCR directly from cell colonies (colony pcr) to check Positive clones.

So Checking DNA isolation success, using PCR is a bad idea
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