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.
Claims12
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.
Dependencies are inferred from the claim text.