Zymo Methylation Gold DNA bisulphite conversion

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lah31
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Zymo Methylation Gold DNA bisulphite conversion

Post by lah31 » Sat Feb 27, 2010 1:49 pm

Hi

I have been investigating DNA methylation and am having trouble with the Zymo methylation gold kit. I have used both the optimal amount of input DNA (500ng) and the maximum amount of input DNA (2ug) for a methylated positive control and a non unmethylated negative control. I have followed the protocol exactly and eluted in 10uL. When I run the input and eluted product on a 2% agarose gel, there is DNA present in the input samples but only a band present in the positive control at 2ug. I don't understand where the rest of the DNA has gone.

I was wondering if anyone had used the kit and experienced any similar problems and whether people knew any modifications to the protocol to improve the DNA recovery. Any help would be greatly appreciated becasue I am in a 'hate' science mood at the moment.

Thanks

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