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Measure nucleotides


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#1 Neumark

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Posted 22 April 2009 - 01:21 AM

Hi,

Is there a method to quantify the levels of A, G, C and T in a sample?

I guess like HPLC would work, but I do not know if it is sensitive enough and if I could distinguish between the different nucleotides. Also I would like to detect them when they are monophosphated.

I talked with a chemist at the hospital, but he said that their method was not sensitive enough and he had no time to improve it.

Is there a known lab that does these detections? Preferably in Europe.

Thanks

#2 Carlton H

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Posted 23 April 2009 - 03:30 PM

If you need a high degree of accuracy, my recommendations would just be to sequence it. That wouldn't really help the quantification of monophosphated nucleotides, but it would give you an effective quantification of the nucleotides. There have to be labs in europe that do it, although I'm from the US and not familiar with commercial sequencing labs across the pond.

-Carlton
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#3 Neumark

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Posted 27 April 2009 - 07:58 AM

Yes, but I am trying to determine the amount an incorporated nucleotide-analogue drug and it is not distinguishable compared to the original nucleotide when sequencing it.

However, if one could measure it after DNA digestion I could get some idea on how much is incorporated.

Regards

#4 Carlton H

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Posted 08 May 2009 - 07:03 AM

I don't know why I didn't think of this earlier, and I'm not an expert on it so I'm not 100% sure it would be sensitive enough, but have you thought about LC/MS?

-Carlton

Edited by Carlton H, 08 May 2009 - 07:04 AM.

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