Novel Ligase Activity
AssigneeNew England Biolabs, Inc.
InventorsGregory Lohman, Thomas C. Evans
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.
Claims15
A composition, comprising an RNA splint ligase and at least one polynucleotide having a length of at least 8 nucleotides in a buffer.
A method for ligating single stranded polynucleotide fragments, comprising:
- (a) combining at least two single stranded polynucleotide fragments having complementary regions at a splice junction to an RNA splint, and an RNA splint ligase;
- and (b) permitting the at least two single stranded polynucleotides to ligate to form a single polynucleotide.
A method for analyzing mRNA for its splicing history, comprising:
- identifying splice junctions, splicing variants or mutations at the splice junction by combining single stranded polynucleotides with the mRNA and an RNA splint ligase.
A method for detecting RNA sequences comprising:
- (a) annealing polynucleotides having regions that are complementary at a ligation junction to a splint RNA;
- (b) ligating the polynucleotides using an RNA splint ligase;
- (c) amplifying the ligation product;
- and
- (d) detecting and optionally quantifying the amplification product.
Dependencies are inferred from the claim text.