Back to results
US 10,597,650 B2Granted
Ligase activity
AssigneeNew England Biolabs, Inc.
InventorsGregory Lohman, Thomas C. Evans, Larry A. McReynolds
Life of this patent
Estimating the term…
Abstract
Compositions and methods are provided for ligating polynucleotides having a length that is greater than 8 nucleotides on an RNA splint. The ligation reaction provides consistent results in high or low ATP concentrations. The reaction can occur rapidly and is generally at least 10 fold more efficient than T4DNA ligase under optimal conditions for T4DNA ligase and the reaction time is less than 6 hours for example, less than 1 hour.
Claims14
A method for ligating DNA polynucleotide sequences, comprising:
- (a) obtaining RNA from a sample that comprises cells or a bodily fluid to produce an RNA sample comprising naturally-occurring RNAs;
- (b) hybridizing at least two DNA polynucleotide sequences to a naturally-occurring RNA in the RNA sample;
- (c) ligating the at least two DNA polynucleotide sequences to one another using a ligase that has an amino acid sequence that is at least 90% identical to the Chlorella virus PBCV-1 ligase of SEQ ID NO:1:
- and
- (d) detecting the product of step (c).
2 more elements
Dependencies are inferred from the claim text.