restriction map - gel

[spinkster]

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]

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]

Cheers for your help cat