HomeFAQsI have run my DNA digested with CviAII in a gel and I cannot see the digested fragment that I was expecting. Has the enzyme degraded my substrate DNA?
FAQ: I have run my DNA digested with CviAII in a gel and I cannot see the digested fragment that I was expecting. Has the enzyme degraded my substrate DNA?
No. Under conditions of excess enzyme, CviAII can bind the substrate DNA quite tightly, so when the DNA is run under standard conditions the DNA and the enzyme bound to it will shift and appear as large DNA fragments in a gel. When running DNA digested with CviAII in a gel the addition of a loading buffer containing 1%SDS is recommended. The SDS will help dissociate the enzyme from the substrate DNA.
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