Abstract: Disclosed herein is a polymerase chain reaction (PCR)-based method for detecting an insertion/deletion variant, as compared with a reference sequence, in a selected region of a target gene in a sample. The target gene has a template strand and a coding strand complementary to the template strand. The method uses a primer set that includes a blocking primer, and a forward primer. The blocking primer and the forward primer are partially overlapping and have different melting temperatures, and the 3?-end of the blocking primer is modified to prevent the extension of the blocking primer during the PCR. Accordingly, under specific PCR conditions, the presence of the PCR product is indicative of the presence of an insertion/deletion mutation in the selected region, and the absence of the PCR product is indicative of the absence of an insertion/deletion mutation in the selected region.

Abstract: Disclosed herein is a self-competitive primer for preferentially amplifying a sample nucleic acid based on whether a selected region thereof has a nucleotide variation, in comparison with a selected region of a reference nucleic acid. The self-competitive primer includes a 5?-competing domain and a 3?-elongating domain. Sequences of the 5?-competing domain and the 3?-elongating domain are complementary to a first region and a second region of the reference nucleic acid, respectively. The first region includes the selected region and at least one nucleotide residue immediately downstream of the selected region of the reference nucleic acid. The second region is located downstream of the selected region of the reference nucleic acid and does not comprise the selected region of the reference nucleic acid, and the first region and the second region overlap by at least one nucleotide.

Abstract: Disclosed herein is a self-competitive primer for preferentially amplifying a sample nucleic acid based on whether a selected region thereof has a nucleotide variation, in comparison with a selected region of a reference nucleic acid. The self-competitive primer includes a 5?-competing domain and a 3?-elongating domain. Sequences of the 5?-competing domain and the 3?-elongating domain are complementary to a first region and a second region of the reference nucleic acid, respectively. The first region includes the selected region and at least one nucleotide residue immediately downstream of the selected region of the reference nucleic acid. The second region is located downstream of the selected region of the reference nucleic acid and does not comprise the selected region of the reference nucleic acid, and the first region and the second region overlap by at least one nucleotide.

Abstract: Disclosed herein is a polymerase chain reaction (PCR)-based method for detecting an insertion/deletion variant, as compared with a reference sequence, in a selected region of a target gene in a sample. The target gene has a template strand and a coding strand complementary to the template strand. The method uses a primer set that includes a blocking primer, and a forward primer. The blocking primer and the forward primer are partially overlapping and have different melting temperatures, and the 3?-end of the blocking primer is modified to prevent the extension of the blocking primer during the PCR. Accordingly, under specific PCR conditions, the presence of the PCR product is indicative of the presence of an insertion/deletion mutation in the selected region, and the absence of the PCR product is indicative of the absence of an insertion/deletion mutation in the selected region.