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Position: Home > Articles > Optimization of RAPD-PCR reaction system and DNA extraction from Ligularia fischeri Agricultural Science Journal of Yanbian University 2011,33 (1) 60-63

蹄叶橐吾基因组DNA提取及RAPD-PCR体系的优化

作  者:
车彩凤;李烨;彭浩;吕龙石
单  位:
延边大学农学院
关键词:
蹄叶橐吾;基因组DNA;RAPD;体系优化
摘  要:
为利用RAPD技术探讨长白山区橐吾属植物的遗传多样性,采用改良的CTAB法成功提取蹄叶橐吾的基因组DNA,并建立橐吾属植物RAPD-PCR反应的最佳体系.最佳体系容积为20μL,其中包括模板DNA20ng,引物10pmol/L,dNTP 300μmol/L,MgCl21.5mmol/L,TaqDNA聚合酶1.5U,不足部分用DW补充.反应程序为94℃预变性5min;94℃变性1min,36℃退火1min,72℃延伸1min 30s,循环40次,最终72℃延伸10min.
译  名:
Optimization of RAPD-PCR reaction system and DNA extraction from Ligularia fischeri
作  者:
CHE Cai-feng,LI Ye,PENG Hao,LV Long-shi*(Agricultual College of Yanbian University,Yanji Jilin 133002,China)
关键词:
Ligularia;genome DNA;RAPD;optimization
摘  要:
To investigation genetic diversity of Ligularia fischeri in Changbai Mountain by molecular biology techniques.The genomic DNA from the Ligularia genus was successfully extracted by using the developed CTAB method and a steady system of PCR reaction for Ligularia fischeri was established.The best RAPD-PCR reaction system optimized is: total volume 20 μL,containing template DNA 20 ng,primer 10 pmol,dNTPs 300 μmol/L,MgCl2 1.5 mmol/L,DNA polymerase 1.5 U,left complement with ddH2O.PCR amplification condition is: pre-denaturation at 94 ℃for 5 min,Denaturation at 94 ℃ for 1 min,annealing at 36 ℃for 1 min,extension at 72 ℃for 1.5 min,cycle 40 times.Final extension at 72 ℃for 10 min.

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