PCR [polymerase chain reaction]

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rkreddy.d
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PCR [polymerase chain reaction]

Post by rkreddy.d » Mon Oct 16, 2006 11:42 am

Hello to community
Is there any efficient method other than classical method to extract mitochondrial DNA and chloroplast DNA from plant seedlings especially from paddy seedlings?

In my lab we used to set PCR without extracting the DNA. We are taking direct plant material using small micropipette tips and sterile toothpicks. With this method we are getting very good reactions. In this case we set primers for chromosomal DNA. But when we do the same with mitochondrial DNA we are unable to get the reaction.

Can any one suggest to solve the problem? :idea:

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LilKim
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Post by LilKim » Mon Oct 16, 2006 5:17 pm

You can use the toothpick,micropipette "mash" techniquefor simple genotyping stuff.

However for you to be able to use the same technique for mDNA you'd have to mash cell wall, cell membrane, mitochondiial inner & outer membrane to lyse ... to realease the mDNA.

Theoretically the mash technique should work... but i'm guessing the the probability of this 'working out' is not so great. So, to make life easier... i think you should comsider using a DNA extraction kit.

good luck!

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