caa a22 4500 378897926 CHVBK 20180305123510.0 cr unu---uuuuu 161128e20040216xx s 000 0 eng 10.1515/CCLM.2004.004 doi (NATIONALLICENCE)gruyter-10.1515/CCLM.2004.004 Evaluation of a new efficient procedure for single-nucleotide polymorphism genotyping: tetra-primer amplification refractory mutation system-polymerase chain reaction [Elektronische Daten] [Naoko Okayama, Kozue Fujimura, Junji Nakamura, Yutaka Suehiro, Yuichiro Hamanaka, Yuji Hinoda] Tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) is a new efficient method for single-nucleotide polymorphism (SNP) genotyping. To determine the optimal conditions for ARMS-PCR we attempted to genotype ten SNPs. DNA was extracted from the peripheral blood of 168 unrelated healthy Japanese volunteers. Two problems inhibited uniform efficiency of the amplification of three bands. The first problem was the lower amplification efficiency of the shorter and allele-specific products compared with the largest product. This phenomenon was overcome by increasing the relative concentration of the inner primers. The second problem was non-specific amplification of the shorter products. To reduce the amplification of these nonspecific bands, adjusting any one of the following PCR conditions was effective: i) reducing the ratio of the inner primer concentration relative to that of the outer primers; ii) increasing the annealing temperature for the initial 5-10 cycles; iii) hot start PCR. With these procedures all ten of the SNPs were successfully genotyped. Our present data may be useful in the further application of tetra-primer ARMS-PCR to SNP genotyping. Copyright © 2004 by Walter de Gruyter GmbH & Co. KG Medical equipment & techniques nationallicence Medical diagnosis nationallicence Diseases & disorders nationallicence Okayama Naoko aut Fujimura Kozue aut Nakamura Junji aut Suehiro Yutaka aut Hamanaka Yuichiro aut Hinoda Yuji aut Clinical Chemistry and Laboratory Medicine Walter de Gruyter 42/1(2004-02-16), 13-16 1434-6621 42:1<13 2004 42 cclm https://doi.org/10.1515/CCLM.2004.004 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/CCLM.2004.004 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Okayama Naoko aut NATIONALLICENCE 700 1- Fujimura Kozue aut NATIONALLICENCE 700 1- Nakamura Junji aut NATIONALLICENCE 700 1- Suehiro Yutaka aut NATIONALLICENCE 700 1- Hamanaka Yuichiro aut NATIONALLICENCE 700 1- Hinoda Yuji aut NATIONALLICENCE 773 0- Clinical Chemistry and Laboratory Medicine Walter de Gruyter 42/1(2004-02-16), 13-16 1434-6621 42:1<13 2004 42 cclm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter