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Quick method for DNA extraction using lysis

DNA isolation PURE DNA isolation Bacterial DNA isolation

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4 replies to this topic

#1 Ameya P

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Posted 09 May 2014 - 11:01 PM

Recently I read in brief about a method of bacterial DNA extraction where in the researchers were using lysis to break open the cells and then somehow managed to concentrate PCR inhibitors in the same tube itself, before using the lysis solution as a template for their PCR. They called it the PURE method DNA extraction. But I have not been able to find any protocol which manages to do something like this. 


Have you heard about such a protocol/ used it yourself. Could you please forward me some details. 

Thanks, 
Ameya 

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#2 BioMiha

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Posted 06 July 2014 - 12:36 PM

We use something similar to this: QuickExtract™ DNA Extraction Solution from Epibio http://www.epibio.co...action-solution


Edited by BioMiha, 06 July 2014 - 12:36 PM.


#3 phage434

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Posted 06 July 2014 - 12:45 PM

Most bacteria can be added directly to a PCR reaction, and the initial denaturation extended for 2-3 minutes to lyse the bacteria. The most common problem with this technqiue is people adding too many bacteria. I'd recommend using a pipet tip to add very small amounts of a colony to 50 ul of water, mixint, then using the same tip, adding 1 ul of the water to the pcr reaction. You can use the same tip to spot a colony on an index plate for later use. The expensive reagents don't do much.



#4 BioMiha

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Posted 06 July 2014 - 12:53 PM

Most bacteria can be added directly to a PCR reaction, and the initial denaturation extended for 2-3 minutes to lyse the bacteria. The most common problem with this technqiue is people adding too many bacteria. I'd recommend using a pipet tip to add very small amounts of a colony to 50 ul of water, mixint, then using the same tip, adding 1 ul of the water to the pcr reaction. You can use the same tip to spot a colony on an index plate for later use. The expensive reagents don't do much.

I agree with phage434. It's called colony PCR. You can find protocols on the web. 



#5 Ameya P

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Posted 11 July 2014 - 12:04 AM

Dear Miha & Phage, 

 

Thank you for your replies. 

 

Colony PCRs are good when you have cultures of the sample. I am looking at doing PCR directly on blood samples without spending too much time/ resources (spin columns, centrifuges etc) on DNA extraction. That is why, I was looking for a simple extraction technique and then came across this PURE method DNA extraction. But have not been able to find the protocol for it. 


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