Stable composition comprising a nuclease and a phosphatase
AssigneeUSB Corporation
InventorsRobert B. Moffett, Jeannine Muller-Greven
Life of this patent
Estimating the term…
Abstract
A composition containing a nuclease, preferably Exonuclease I, and a phosphatase, preferably Shrimp Alkaline Phosphatase, wherein the enzymes are combined in a single composition yet each enzyme retains significant functional activity over time. Combining Exonuclease I and Shrimp Alkaline Phosphatase into one composition allows simplified processing of amplified DNA to degrade residual primers and nucleotide triphosphates thereby facilitating subsequent DNA analysis.
Claims37
A method of degrading preselected nucleic acids present in a sample of material, the method comprising the step of contacting said sample with a composition comprising a nuclease and a phosphatase, said nuclease degrading at least a portion of said prescribed nucleic acids present in said sample of material, wherein said nuclease is present in said composition in a concentration of at least 0.01 units of nuclease per microliter of composition, and said phosphatase retains at least 50% of its functional activity when said composition is stored at −20° C. for 24 hours.
A method comprising the steps of:
- (a) amplifying DNA by a PCR amplification reaction to provide an amplification reaction product comprising amplified DNA;
- (b) contacting said amplification reaction product with a composition comprising a nuclease and a phosphatase, wherein, prior to being contacted with said amplification reaction product, said composition is substantially free from the presence of amplified deoxyribonucleic, said nuclease is present in said composition in a concentration of at least 0.01 units of nuclease per microliter of composition, and said phosphatase retains at least 50% of its functional activity when said composition is stored at −20° C. for 24 hours;
- and (c) performing a single nucleotide polymorphism analysis of said amplification reaction product following said contacting step to determine a single nucleotide difference in said amplified DNA.
Dependencies are inferred from the claim text.