Method of nucleotide identification using an off-switch through proofreading 3' exonuclease-resistant modified primers by polymerases with 3' exonuclease activity

摘要

Methodology for a high fidelity polymerase-mediated primer extension for use in genotyping is provided. The primer extensions are carried out with polymerase having 3′ to 5′ exonuclease activity and primers with their 3′ modification of being exonuclease-resistant. The primers are designed to have their 3′ termini: 1. complementary to nucleotide to be analyzed; and 2. subject to be proofread by the 3′ to 5′ exonuclease of the high fidelity polymerases.