supercoiled DNA migration in agrose ?

Victor Jongeneel vjongene at isrec-sun1.unil.ch
Mon Dec 28 01:57:42 EST 1992


chai_z at wehi.edu.au wrote:

: Hi,

: 	Several batches of M13 RF DNA of mine have been cut by EcoRI or SmaI,
: etc. and then electrophoresized in agrose gel. Uncut DNA (supercoiled) of some
: batches migrated faster than the cut one (linear), while the others slower.
: Most likely, some batches of my DNA were nicked (open circular). Could any one
: tell me the correct agrose pattern of supercoiled, open circular and linear DNA?
: or tell me where to refer to?

: Thanks in advance.

: Zhonglin Chai 


In most agarose gels (but _not_ always), supercoiled circular DNA
migrates fastest, followed by linear and nicked circles.  Linear does
not always separate from supercoiled, nicked circles usually do.  This
does of course not exclude the presence of miniphage (see other
response in thread).  The best way to find out is to start
with freshly purified (CsCl/EtBr) supercoiled DNA and treat it so as
to make linears and nicked circles.

Cheers,


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C. Victor JONGENEEL                       vjongene at isrecmail.unil.ch
Ludwig Institute for Cancer Research
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