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Information about the use of DNA diluted in Real Time PCR

DNA Dilutions Real Time PCR

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6 replies to this topic

#1 rmmoniz

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Posted 19 September 2013 - 07:29 AM

Hi, 

For the removal os PCR inhibitors for Real Time Detection we should dilute the DNA sample 1:10 fold, consequently we are reducing the amount of DNA available for amplification, this fact cannot give false- negative results? 

Thanks



#2 bob1

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Posted 19 September 2013 - 03:33 PM

Why not?



#3 rmmoniz

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Posted 20 September 2013 - 07:21 AM

So for PCR inhibitors removal, what do you recomend to use in daily diagnosis to prevent false negative results?



#4 bob1

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Posted 20 September 2013 - 07:48 PM

There isn't anything that I am aware of.  The false negatives won't be entirely removed by diluting 1:10, you could also try 1:100, 1:1000 etc.



#5 rmmoniz

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Posted 23 September 2013 - 04:32 AM

But the dilutions that you recommend wont increase the chances of the sample give a false negative result? Mainly in the cases of very minimum loads of DNA/ RNA target?



#6 bob1

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Posted 25 September 2013 - 01:21 AM

Potentially, though if you are doing qPCR with good primers, then the reaction should be close to 100% efficient, so detection should work from very low abundance targets.



#7 rmmoniz

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Posted 25 September 2013 - 07:37 AM

Ok, thank you very much laugh.png  







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