restriction map - gel

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spinkster
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restriction map - gel

Post by spinkster » Tue Mar 18, 2008 2:32 pm

Hi guys,

I have done a restriction map gel and got what I expected, but my uncut pUCPH seemed to seperate into multiple bands.
(I was only expecting 1 band as surley no restriction sites used therefore surely it should all be one size)

does anybody know why this is?

any help would be much apprieciated

Matt

Cat
King Cobra
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Post by Cat » Tue Mar 18, 2008 3:19 pm

To get a single band, you need to cut your vector with a single cut enzyme. Uncut plasmids can be a mixture of topologically different forms (nicked, linear, covalently bonded, single stranded circular, and supercoiled) and none of them will migrate as expected of the cut linear vector.

spinkster
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Re: restriction map - gel

Post by spinkster » Wed Mar 19, 2008 9:46 am

Cheers for your help cat :)

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