基于4300 DNA分析系统的茶树SSR发掘方法优化与建立

doi:10.13305/j.cnki.jts.2014.05.009

Abstract

Abstract: Simple sequence repeats (SSR) play an important role in genetic and breeding research for tea plant [Camellia sinensis (L.) O. Kuntze]. 4300 DNA Analysis System is high-throughput, accurate and sensitive in detecting the signals, it has been used in molecular markers techniques in many species, but its application in tea plant has not been reported up to now. In this study, the main parameters that affect tea SSR-PCR have been optimized and verified via single factor experiment and L₉(3⁴) orthogonal test. A suitable SSR-PCR reaction system of the 4300 DNA Analysis System for the tea plant genetic research was obtained: 1.0 μL DNA（25 ng·μL^-1）, 0.2 μL MF-F, 0.2 μL R and 0.40 μL IR-MF, 0.8 μL of dNTPs (25 mmol·L^-1), 1.0 μL 10×Buffer (Mg²⁺), 0.1 μL Ex-Taq polymerase (5 U·μL^-1), adding sterile water to a total volume of 10 μL. Concentration of all primers is 1 μmol·L^-1. Besides, in this research, polyacrylamide gel solution (acry:bis is 29:1, concentration is 6.5%) self-made is detected to be alternative to the gel solution provided by 4300 DNA analysis system for detecting SSR loci.

Key words: tea plant, 4300 DNA Analysis System, SSR-PCR

CLC Number:

S571.1

ZHANG Jieru, WEI Chaoling. Establishment a SSR-PCR System of Tea Plant [Camellia sinensis (L.) O. Kuntze] Based on the 4300 DNA Analysis System[J]. Journal of Tea Science, 2014, 34(5): 481-488.

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Establishment a SSR-PCR System of Tea Plant [Camellia sinensis (L.) O. Kuntze] Based on the 4300 DNA Analysis System

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