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How much product PCR??


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#1 RedRosa

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Posted 20 October 2009 - 04:43 AM

Hi all,

I have a question: how much should I take PCR product for electrophoresis and what about precentage of the gel?

If DNA after modyfication is so fragile. If I took not enough PCR product I could not see it, could I?

#2 almost a doctor

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Posted 20 October 2009 - 08:31 AM

Hi all,

I have a question: how much should I take PCR product for electrophoresis and what about precentage of the gel?

If DNA after modyfication is so fragile. If I took not enough PCR product I could not see it, could I?



I was told long time ago that if I cant see 1/10 of my PCR on a gel, it hasnt worked, although I guess that's probably change now with more sensitive detection systems. Also depends what you are looking for and what you want to do with the product.
As for percentage of gel that'll depend on the size of your product, I generally run 1% agarose but more for convenience than anything else.

What volume PCR do you have, and what do you want to do with it?

I dont understand your second question, sorry. What you mean by DNA modification?

#3 RedRosa

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Posted 22 October 2009 - 09:07 AM

I want to sequence my PCR product. We talk about methylation so I use " modyfication" maby it is bad term, better conversion?

I am not sure if I took enough PCR product for electrophoresis, maby that is the reason that I do not see anything

That is why I asked how much should I take it?

Edited by RedRosa, 22 October 2009 - 09:09 AM.


#4 Adrian K

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Posted 22 October 2009 - 03:36 PM

I never done methylation so i not be sure, but for sequencing purpose, I will load all my products into the gel, to gel purified it before send to sequencing. Tried a gene with 2.5k size, no problem.
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