摘要
利用聚合酶链反应(Polymerasechinreaction,PCR)技术,特异性扩增沙门菌HI基因的269bp片段。8株标准阳性菌和3株阳性菌检测结果完全相符,并对30株分离的疑似沙门氏菌进行了检测。采用50μl体系,引物为自行设计各206割寡聚核苷酸,各1μM;dNTPs各100μM;Taq酶2U。二温段水浴扩增35循环,条件为预变性97℃7min,变性94℃60s,复性和延伸60℃60s。
HI priners which targeted a 269 bp region of HI gene inSalmonella,were used in the polymerase chain reaction(PCR). The standard PCR was done i 50 μl volume. It contained 1μM of eachrinier,2.5mM Mgcl 2,200μM dNTPs,104 ~ 108 copies of templates DNA. The amplification was performed by mannual operation between twotemperature control scations . Temperature cycle was designde in former denaturation at 97℃ 7min,35 cycles with denaturation 94℃ 60 s anneating and extension at 60℃ 60s,and last estension at 72℃ 7min.
出处
《黑龙江畜牧兽医》
CAS
北大核心
1997年第2期7-9,共3页
Heilongjiang Animal Science And veterinary Medicine