Establishment and optimization of AFLP reaction system for Akebia trifoliata
- Author:
Zhe WANG
1
Author Information
1. College of Life Science
- Publication Type:Journal Article
- Keywords:
AFLP;
Akebia trifoliata (thunb.) koidz;
Optimization
- From:
Chinese Traditional and Herbal Drugs
2010;41(12):2074-2078
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish and optimize the AFLP reaction system for genetic diversity analysis on Akebia trifoliata. Methods: The leaves of A. trifoliata were used to analyze the influencing factors of the ligation, the pre-amplification, and the selective amplification of AFLP, so that the optimal AFLP reaction system could be built up. Results: The optimal AFLP reaction system has been set up: added 10 pmo1 of EcoR I adapter, 100 pmol of Mse I adapter, and 2 U of T4 DNA ligase in the ligation mixture; The pre-amplification mixture (20 μL) contained 2. 0 μL of ligation products, 100 ng of each pre-selective primer, E-0 and M-0, 0. 250 mmol/L of dNTPs, 1. 875 mmol/L of Mg2+, 2 U of Taq DNA polymerase; the selective amplification mixture (20 μL) contained 5 μL of 1 : 100 diluted pre-amplification products, 50 ng of each selective primer, E3, M3, 0. 250 mmol/L of dNTPs, 1. 875 mmol/L of Mg2+, 1. 5 U of Taq DNA polymerase. Conclusion: The study on A. trifoliata could establish and optimize the AFLP reaction system which can be used in the research of genetic diversity of the species in future.
