广东农业科学
廣東農業科學
엄동농업과학
GUANGDONG AGRICULTURAL SCIENCES
2010年
1期
120-123
,共4页
野葛%粉葛%ISSR-PCR%正交优化
野葛%粉葛%ISSR-PCR%正交優化
야갈%분갈%ISSR-PCR%정교우화
Pueraria lobata (Willd) Ohwi%Pueraria thomsonii Benth%ISSR-PCR%orthogonal optimization
利用正交试验设计法L_9(3~4),从引物浓度、TaqDNA聚合酶浓度、Mg~(2+)浓度和dNTP浓度4种因素3个水平,对葛资源ISSR-PCR反应体系进行优化分析,并在此基础上对PCR反应的退火温度进行梯度筛选.结果表明,25μL最佳的葛资源ISSR-PCR反应体系是:1×PCR buffer、0.20 mmol/L dNTP、0.4 μmol/L引物、2.5 mmol/L Mg~(2+)和0.5 U TaqDNA聚合酶,引物UBC809的最佳退火温度为57.9℃.
利用正交試驗設計法L_9(3~4),從引物濃度、TaqDNA聚閤酶濃度、Mg~(2+)濃度和dNTP濃度4種因素3箇水平,對葛資源ISSR-PCR反應體繫進行優化分析,併在此基礎上對PCR反應的退火溫度進行梯度篩選.結果錶明,25μL最佳的葛資源ISSR-PCR反應體繫是:1×PCR buffer、0.20 mmol/L dNTP、0.4 μmol/L引物、2.5 mmol/L Mg~(2+)和0.5 U TaqDNA聚閤酶,引物UBC809的最佳退火溫度為57.9℃.
이용정교시험설계법L_9(3~4),종인물농도、TaqDNA취합매농도、Mg~(2+)농도화dNTP농도4충인소3개수평,대갈자원ISSR-PCR반응체계진행우화분석,병재차기출상대PCR반응적퇴화온도진행제도사선.결과표명,25μL최가적갈자원ISSR-PCR반응체계시:1×PCR buffer、0.20 mmol/L dNTP、0.4 μmol/L인물、2.5 mmol/L Mg~(2+)화0.5 U TaqDNA취합매,인물UBC809적최가퇴화온도위57.9℃.
In this study, orthogonal experimental design method L_9 (3~4)was used, from the concentration of primers, Taq DNA polymerase concentrations, Mg~(2+) concentration and dNTP concentrations of four kinds of factors in three levels, ISSR-PCR reaction system of Pueraria was analysed. On this basis, the annealing temperature were proposed by gradient PCR. The results showed that, 25 μL ueraria resources ISSR-PCR reaction system was: 1×PCR buffer, 0.20 mmol/L dNTP, 0.4 μmol/L primer, 2.5 mmol/L Mg~(2+)and 0.5 U Taq DNA polymerase, the optimal annealing temperature of primer UBCS09 was 57.9℃.
