The effects of several amplification conditions on RAPD amplification for Alsophila spinulosa
LI Xue-yan1, WANG Ting2, SU Ying-juan2
1. Department of Biotechnology of Zhongshan College, University of Electronic Science and Technology of China, Zhongshan 528402; 2. School of Life Sciences, Zhongshan University, Guangzhou 510275, China
To get the optial amplification conditions for Alsophila spinulosa,the effects of the purified methods of DNA templates and the concentrations of DNA templates,Taq DNA polymerase,magesium,primer and dNTP are tested.The results suggest:the purified templates by glass milk or proteinase K are brighter,clearer and more repeatable than the unpurified templates or precipitated templates by ethanol;the concentration of DNA templates,Taq DNA polymerase,magesium,primer and dNTP will influence the productions of RAPD.The optimal amplification conditions for Alsphila spinulosa is acquired through the experiments:the concentration of DNA templates is 70ng,the concentration of magnesium is 2.5 mmol·L-1,dNTP is 0.2 mmol·L-1,primer is 3×104 mL.The optical amplification program is also obtained:94·200s,1 cycle;94℃ 60s,36℃ 60s;72℃ 120s,40 cycle;72℃ 600s,1 cycle;stored in 4℃.
LI Xue-yan,WANG Ting,SU Ying-juan. The effects of several amplification conditions on RAPD amplification for Alsophila spinulosa[J]. , 2003, 22(2): 120-123.
[6] Anne R K, Les J S. 1995. High-GC primer are useful in RAPD analysis in fungi[J]. Curr Genet, 28:384～389.
[7] Benter T, Papadopoulos S, Pape M, et al. 1995.Optimization and reproducibility of random amplified polymorphic DNA in human[J]. Anal Biochem,230(1):92～100.
[8] Devos K M, Gale M D. 1992. The use of random amplified polymorphic DNA markers in wheat[J]. TAG, 84:567～572.
[9] Ellsworth D L, Rittenhouse K D, Honeycutt R L. 1993.Aritifactual variation in randomly amplified polymorphic DNA banding patterns[J]. Biotechniques, 14:214～217.
[10] Thormann C E, Ferreira M E, Camargo L E A, et al. 1994.Comparison of RFLP and RAPD markers to estimating genetic relationships within and among cruciferous species[J]. Theor Appl Genet, 88:973～980.
[11] Yong H P, Russell J K. 1994. Effect of concentration of MgCl2 on random amplified DNA polymorphism [J].Biotechniques, 16(4):652～656.
