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- Title
Establishment of the Optimized AFLP Analysis System for Alnus.
- Authors
Longbing RAO; Zhichao ZHANG; Hanbo YANG; Hongying GUO; Hongping DUAN; Yitai CHEN
- Abstract
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 tme ad amplification system that influenced te results of the AFLP. The results suggested an amount of DNA template of 300 ng; reaction time for enzyme digestion of 5 h; ligaton 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 pi) for Alnus was determined as follows : DNA template 300 ng, 10 X Taq DNA polymerase Buffer (containing Mg2 + ) 2. 0 pl, dNTPs 0.3 pl, primers 1.0 pl, Taq DNA polymerase 0. 6 U, and ddH20 in balance. Seven pairs of primer combinations were selected with many clear bads and high polymorphism. By using the AFLP reaction system, primers suitable for Alnus were obtained. Those results provided certain bais for further molecular studies on Alnus using AFLP markers.
- Subjects
AMPLIFIED fragment length polymorphism; ALNUS glutinosa; GENETIC polymorphisms in plants; PLANT enzymes; ALDER
- Publication
Agricultural Biotechnology (2164-4993), 2015, Vol 4, Issue 5, p37
- ISSN
2164-4993
- Publication type
Article