Abstract: Provided herein are methods for ligation of polynucleotides containing modified ligation components, particularly modified ligase cofactors, modified acceptors and modified donors. The methods readily applied to ligation-based assays for detection of a nucleic acid sequence where the use of the modified cofactor improves discrimination between matched and mismatched templates. Furthermore, the use of the modified ligation components reduces or eliminates the ligation in the absence of nucleic acid template.

Abstract: Provided herein are methods and compositions for nucleic acid replication. These methods involve the use of 3?-substituted nucleoside 5?-triphosphates or 3?-substituted terminated primers in nucleic acid replication reactions. In certain aspects, the methods are accomplished by use of 3?-substituted NTPs and/or 3?-substituted terminated primers which provide utility in nucleic acid replication. In preferred embodiments, the NTPs and/or primers are substituted at the 3?-position with particular heat labile chemical groups such as ethers, esters or carbonate esters.

Abstract: The present invention provides methods and compositions for nucleic acid amplification. These methods involve the use of oligonucleotide primers in temperature dependent nucleic acid amplification reactions. In certain aspects, the methods are accomplished by use of certain modified oligonucleotide primers which provide utility in nucleic acid amplification. In preferred embodiments, the oligonucleotide primers are modified with particular chemical groups such as esters.