Methods and compositions for improving efficiency of nucleic acids amplification reactions

摘要

The present invention provides methods and compositions for improving the efficiency of nucleic acid amplification reactions. The invention encompasses hybrid polymerases that show increased processivity over wild type polymerases as well as decreased exonuclease activity. The invention also encompasses methods, compositions and kits for conducting nucleic acid synthesis and amplification reactions in which non-specific amplification of primers is reduced.

主权项

1. A reaction mixture comprising:
a hybrid polymerase, wherein said polymerase comprises a polymerase domain and a heterologous DNA binding domain; an additive selected from the group consisting of sarcosine and heparin, in a sufficient amount to improve efficiency of the amplification reaction by at least 10% compared to a reaction mixture lacking the additive, and an antibody that is specific for the hybrid polymerase, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the variable regions comprise complementary determining regions (CDRs), wherein:
(a) the heavy chain variable region CDRs comprise SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24, and the light chain variable region CDRs comprise SEQ ID NO:25, SEQ ID NO:26, and SEQ ID NO:27; or(b) the heavy chain variable region CDRs comprise SEQ ID NO:28, SEQ ID NO:29, and SEQ ID NO:30, and the light chain variable region CDRs comprise SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33; or(c) the heavy chain variable region CDRs comprise SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:36, and the light chain variable region CDRs comprise SEQ ID NO:37, SEQ ID NO:38, and SEQ ID NO:39.