Establishment of the Optimized AFLP Analysis System for Alnus  

作　　者：Longbing RAO Zhichao ZHANG Hanbo YANG Hongying GUO Hongping DUAN Yitai CHEN 

机构地区：[1]Institute of Subtropical Forestry,CAF [2]School of Agriculture and Biological Technic,Yunnan Agricultural University [3]Sichuan Academy of Forestry

出　　处：《Agricultural Biotechnology》2015年第5期37-41,44,共6页农业生物技术（英文版）

基　　金：Supported by National Science&Technology Support Program during the"12th Five-Year"(2012BAD01B0604);Central Public-interest Scientific Institution Basal Research Fund(RISF2013010)

摘　　要：The optimized AFLP analysis system was established for Alnus. A. glutinosa was used to comparatively study and analyze key factors influencing the results of AFLP, including quality and concentration of extracted DNA, enzyme digestion time and amplification system that influenced the results of the AFLP. The results suggested an amount of DNA template of 300 ng; reaction time for enzyme digestion of 5 h; ligation time of 16 h; a 10 times dilution of ligation products for pre-amplification ; and a 20 times dilution of pre-amplification products for selective amplification. The AFLP-PCR reaction system （20 μl） for Alnus was determined as follows ： DNA template 300 ng, 10 x Taq DNA polymerase Buffer （ containing Mg2＋ ） 2.0μl, dNTPs 0. 3 μl, primers 1.0 μl, Taq DNA polymerase 0. 6 U, and ddH2O in balance. Seven pairs of primer combinations were selected with many clear bands and high polymorphism. By using the AFLP reaction system, primers suitable for Alnus were obtained. Those results provided certain basis for further molecular studies on Alnus using AFLP markers.The optimized AFLP analysis system was established for Alnus. A. glutinosa was used to comparatively study and analyze key factors influencing the results of AFLP, including quality and concentration of extracted DNA, enzyme digestion time and amplification system that influenced the results of the AFLP. The results suggested an amount of DNA template of 300 ng; reaction time for enzyme digestion of 5 h; ligation time of 16 h; a 10 times dilution of ligation products for pre-amplification ; and a 20 times dilution of pre-amplification products for selective amplification. The AFLP-PCR reaction system （20 μl） for Alnus was determined as follows ： DNA template 300 ng, 10 x Taq DNA polymerase Buffer （ containing Mg2＋ ） 2.0μl, dNTPs 0. 3 μl, primers 1.0 μl, Taq DNA polymerase 0. 6 U, and ddH2O in balance. Seven pairs of primer combinations were selected with many clear bands and high polymorphism. By using the AFLP reaction system, primers suitable for Alnus were obtained. Those results provided certain basis for further molecular studies on Alnus using AFLP markers.

关 键 词：AFLP A. glutinosa POLYMORPHISM 

分 类 号：S792.14[农业科学—林木遗传育种]