T Nakayama1, M Soma, D Rahmutula, Y Ozawa, K Kanmatsuse. 1. 2nd Department of Internal Medicine, Nihon University School of Medicine, Ooyaguchi-kamimachi 30-1, Itabashi-ku, Tokyo 173-8610, Japan. tnakayam@med.nihon-u.ac.jp
Abstract
BACKGROUND: Isolation of the 5'-flanking region of gene segments adjacent to known sequences is a tedious task. Thermal asymmetric interlaced (TAIL)-PCR developed in 1995 uses nested sequence-specific primers together with a shorter arbitrary degenerate primer so that the relative amplification efficiencies of specific and nonspecific products can be thermally controlled. In the present study, we modified the TAIL-PCR method to isolate the 5'-flanking region of two human genes, human natriuretic peptide receptor type A (NPRA) and type B (NPRB) genes. MATERIAL AND METHODS: We improved upon the original TAIL-PCR method in some aspects: three different series of eight AD primers were used simultaneously; a high denaturing temperature was used; and a Taq DNA polymerase developed for long and accurate PCR was used. RESULTS: We succeeded in isolating the 5'-flanking region of the NPRA and NPRB genes directly from human genomic DNA. CONCLUSION: Our TAIL-PCR method is applicable not only to be human genome but also to the genome of other life forms.
BACKGROUND: Isolation of the 5'-flanking region of gene segments adjacent to known sequences is a tedious task. Thermal asymmetric interlaced (TAIL)-PCR developed in 1995 uses nested sequence-specific primers together with a shorter arbitrary degenerate primer so that the relative amplification efficiencies of specific and nonspecific products can be thermally controlled. In the present study, we modified the TAIL-PCR method to isolate the 5'-flanking region of two human genes, human natriuretic peptide receptor type A (NPRA) and type B (NPRB) genes. MATERIAL AND METHODS: We improved upon the original TAIL-PCR method in some aspects: three different series of eight AD primers were used simultaneously; a high denaturing temperature was used; and a Taq DNA polymerase developed for long and accurate PCR was used. RESULTS: We succeeded in isolating the 5'-flanking region of the NPRA and NPRB genes directly from human genomic DNA. CONCLUSION: Our TAIL-PCR method is applicable not only to be human genome but also to the genome of other life forms.
Authors: T Nakayama; T Nakazato; H Naruse; Z Fu; Z Wang; M Soma; T Hoshino; M Shimodaira; N Aoi Journal: J Hum Hypertens Date: 2017-02-09 Impact factor: 3.012
Authors: Jannik Fonager; Blandine M D Franke-Fayard; John H Adams; Jai Ramesar; Onny Klop; Shahid M Khan; Chris J Janse; Andrew P Waters Journal: BMC Genomics Date: 2011-03-20 Impact factor: 3.969