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Reaction System Optimization and Primer Screening of ISSR Fingerprint Map for Camellia Cultivars(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2014年22期
Page:
84-88
Research Field:
Publishing date:

Info

Title:
Reaction System Optimization and Primer Screening of ISSR Fingerprint Map for Camellia Cultivars
Author(s):
HU Xing-hua; LI Jing-jian; WANG Yan; HUANG Shi-xun
Guangxi Institute of Botany,Guangxi Zhuang Autonomous Region and the Chinese Academy of Sciences,Guilin,Guangxi 541006
Keywords:
Camellia cultivar; molecular fingerprint map; ISSR reaction system optimizing; primer screening
PACS:
S 796
DOI:
-
Abstract:
Taking 30 Camellia cultivars as test materials,the effects of concentrations of Mg2+,dNTP,Primers,Taq DNA polymerase,template DNA,and the annealing temperature for primers on the ISSR-PCR molecular fingerprint map were examined by orthogonal design test and single factor modified.The results showed that the optimal ISSR-PCR conditions in the experiments were as following:in 25 μL reaction system containing Mg2+ 3.0 mmol/L、dNTPs 0.2 mmol/L、primers 0.3 mmol/L、Taq DNA polymerase 0.5 U、template DNA 80 ng and annealing temperature 52.1℃.Base on this optimal reaction system,100 ISSR primers were used to screen for the suitable primers with 1 cultivar samples,of which 12 ISSR primers with high resolution and multiple polymorphic bands were screened.Further amplification of 30 Camellia japonica cultivars using these 12 primers demonstrated that this optimized reaction system had specialties of good repeatability and stability.

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Last Update: 2014-12-12