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Standard PCR Protocol



Standard PCR Protocol

From Molecular Biology Techniques Manual. Protocol written by Ed Rybicki , January 1994, February 2001.

Content

Recommended Reagent Concentrations
* Recommended Reaction Conditions
* Initial Conditions
* Temperature Cycling
* "Hot Start" PCR
* Asymmetric PCR for ssDNA Production
* Detecting Products
* Labelling PCR Products with Digoxigenin
* Cleaning PCR Products
* Sequencing PCR Products
* Cloning PCR Products
* AND ALWAYS REMEMBER:

Last update 17-Mar-2005, Rating of 2 votes.

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By satya on 14-Jun-2007
well iam trying to standardise the pcr protocol from few weeks ...i tried in almost all the ways and combinations....band at specified region came few times but when ever iam trying to reduce the nonspecific bands by increasing anneling temp and all ...total collapse...no amplification at all .....


my question is that whether the pH of the pcr buffer does hav any effect on amplification........
well my desired product is around 300bp...and the pH iam using is 9.0.
well this makes any difference ..if so wat kind of effect does it shows ..........

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By khanfar on 17-Mar-2005
good

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