| 摘要 |
<p>An improved method for performing a primer extension reaction is disclosed, such method including the steps of annealing an oligonucleotide primer to a portion of a template nucleic acid thereby forming a primer-template hybrid; adding primer-extension reagents to the primer-template hybrid for extending the primer; and adding a cosubstrate-enzyme pair to the primer-template hybrid for conducting a pyrophosphate-utilizing reaction, thereby reducing the amount of pyrophosphate present in the reaction. In a particularly preferred embodiment, the cosubstrate-enzyme pair comprises pyrophosphate dependent phosphofructose kinase and fructose-6-phosphate, or UDP glucose pyrophosphorylase and UDP glucose. The invention further includes kits and solutions useful for carrying out the methods of the invention.</p> |
