high sizes on gel - (Dec/09/2006 )

hi

can anyone tell me whats the highest PCR band size that can be seen on agarose gel ?

thnks

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largest DNA fragment that can be resolved?

It depends on your gel box type. A pulse field gel can easily separate out several Mb fragment. Small whole chromosomes.

A normal linear field gradient gel can resolve to about ~15 - 20 kb. though resolution is now in the order of 2000s bp differences... (if memory serves). it is not much good at this level.

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by the way whats the highest size of dna fragment amplified by pcr in one time

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that I have read about ... 50kb
that I have done 8kb
that I have seen done 20kb

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Wouldn't hold your breathe trying to get those sort of sizes done in the one PCR - very difficult. If you are going to attempt it, try a couple of different things but then move on and clone it in 2 kb fragments - not simpler by any means but a lot easier to isolate the products.

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Well there is an art to it. But 2kb is just plain small. 4kb... 4kb isn't hard to do. Clone it as 4kb fragments.

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