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Yasar Yousafzai 03-09-2012 02:01 AM

DNA extraction using TRIzol
 
Hi,
I am using Trizol for extracting genomic DNA from cerebrospinal fluids. I am expecting between 2-5 ng genomic DNA from my sample but in PCR there is no amplification. I have tried several different changes in trizol extraction protocol to get a better yield but nanodrop is not sensitive enough and PCR shows no amplification. I have around 500 human samples lysed in TRIZOL waiting to be extracted but I have failed to optimize the extraction yet.
My questions are:
1. Does Trizol eluted DNA contain any PCR inhibitors? If yes, then how to clean the eluted DNA (I have around 100 samples already extracted)
2. Can I apply the TRIZOL organic phase directly on to a DNA column (DNA micro kit)?
Will be grateful for any tips.
Yasar

cy2chrome 11-15-2012 11:25 PM

Re: DNA extraction using TRIzol
 
Hello,

I am extracting DNA from pelleted CSF cells lysed inTRizol. How many samples have you done so far? What are your average yields? Did you resuspend your DNA in 8mM NaOH as recommended by Invitrogen/Life Sciences? I can't seem to find anyone else who has experience extracting DNA from CSF cells.

Thanks!

--Christine


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