Help!!!Restriction enzyme digestion troubles.

Helen Niederwald helennie at my-deja.com
Fri Nov 10 10:29:21 EST 2000


In article <LAW2-OE458M19VMczi9000001bf at hotmail.com>,
  rodneypettway at hotmail.com ("Rodney Pettway") wrote:
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> I start out with high molecular weight DNA.

What kind? Plasmid? Chromosomal? What source? What purification method?


When I try and carryout my d=
> igestion it seems as though my DNA is being degraded by the enzyme

What else do you expect? It's a restriction enzyme. If you digest
chromosomal DNA you will see a smear.



and I =
> have tried two different enzymes.  When I just add water or buffer or
wat=
> er/buffer to the DNA it is fine i.e. high molecular weight.  what
should =
> I do!!!!!!

Test the enzyme on a standard template. However, most likely nothing is
wrong with your experiment. More likely, your interpretation needs some
refinement.
Regards,
Helen.



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> <HTML><BODY STYLE=3D"font:10pt verdana; border:none;"><DIV>I start
out wi=
> th high molecular weight DNA.&nbsp; When I try and carryout my
digestion =
> it seems as though my DNA is being degraded by the enzyme and I have
trie=
> d two different enzymes.&nbsp; When I just add water or buffer or
water/b=
> uffer to the DNA it is fine i.e. high molecular weight.&nbsp; what
should=
>  I do!!!!!!</DIV></BODY></HTML>
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