两轮多重反转录巢式PCR方法扩增全长GalUR基因.pdfVIP

业生物技术学报 Journal of Agricultural Biotechnology 2005,13 (6): 728~732 ·研究论文· Multiplex Reverse Transcription-nested-double PCR for Generation of Full-length Open Reading Frames of Gene* 1 2 3 YANG Jing-Hui ** Gefu Wang-Pruski Mark Hodges The method of multiplex reverse transcription-nested-double PCR was studied. After the first PCR was prepared with nested primers and cDNA, the PCR mixture was amplified using the same primers. The level of the desired PCR product was in- creased after optimization of reaction conditions with different Mg2+ concentrations, annealing temperatures and annealing duration. The results demonstrated that the multiplex reverse transcription-nested-double PCR method was sensitive enough to amplify full-length genes of strawberry ( cv. Sweat Charlie), in this case . This method is also suitable for amplifica- tion of the full-length gene containing restriction enzyme sites. double- multiplex amplification; nested PCR; full-length gene; 两轮多重反转录巢式PCR 方法扩增全长 基因 1 2 3 杨静慧 ** Gefu Wang-Pruski Mark Hodges (1. 天津农学院园艺系, 天津300381; 2. Nova Scotia 学院植物动物科学系, Truro, NS, 加拿大, B2N 5E3; 3. 加拿大 业食品研究院园艺研究中心, Kentville, NS, 加拿大, B4N