Method for performing multiple enzyme reactions in a single tube
AssigneeNew England Biolabs, Inc.
InventorsRomualdas Vaisvila, Lauren Higgins, Ashley Luck
Life of this patent
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Abstract
Among other things, a method for performing multiple enzyme reactions in a single tube is provided. In some embodiments, the method may comprise producing a reaction mix comprising a thermolabile UDG, an AP lyase and DNA fragments that comprise one or more uracil residues, incubating the reaction mix at a relatively low temperature to cleave fragments at the one or more uracil residues, raising the temperature of the reaction mix to a relatively high temperature to inactivate the thermolabile UDG; and deaminating the fragments, thereby converting any cytosine in the fragments of DNA to uracil.
Claims13
A method for performing multiple enzyme reactions in a single tube, comprising:
- (a) producing a reaction mix comprising a thermolabile uracil-DNA glycosylase (UDG), a DNA-(apurinic or apyrimidinic site) lyase (AP-lyase) and fragments of genomic or plasmid DNA that comprise one or more uracil residues;
- (b) incubating the reaction mix at a temperature of below 40° C. to cleave the genomic or plasmid DNA fragments at the one or more uracil residues;
- (c) raising the temperature of the reaction mix to at least 50° C. for a period of time to inactivate the thermolabile UDG;
- and
- (d) after step (c) deaminating the fragments of genomic or plasmid DNA, thereby converting any cytosine in the fragments of genomic or plasmid DNA to uracil.
13-24. (canceled)
Dependencies are inferred from the claim text.