denaturing vs non denaturing gels
Nick Jacobsen
jacobsen at cf.ac.uk
Tue Jun 10 08:35:16 EST 1997
Hi,
denaturing gels will contain a DNA denaturant e.g. urea which will keep
your DNA fully denatured throughout the gel run. Non-denaturing gels
obviously do not contain urea and promote renaturation of DNA and
secondary structure formation within ssDNA - hence its use in SSCP when
you are looking for single base changes that alter secondary structure.
This leads (in an ideal situation) to differing conformer profiles for
wild type and mutant. (N.B. prior to loading you heat samples to 95 C for
five minutes in formamide loading buffer in order to initially denature
the strands)
hope this helps
Nick.
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