|Table of Contents|

Establishment and Optimization of Chrysanthemum SSR-PCR Reaction System(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2012年13期
Page:
127-131
Research Field:
Publishing date:

Info

Title:
Establishment and Optimization of Chrysanthemum SSR-PCR Reaction System
Author(s):
LI Ya-hui 1; 2; HUANG Cong-lin2; DONG Ran1
1.College of Gardening，Jilin Agricultural University，Changchun，Jilin 130118；
2.Beijing Agro-Biotechnology Research Center，Beijing Academy of Agriculture and Forestry Sciences，Beijing 100097
Keywords:
chrysanthemum; SSR markers; orthogonal design; optimization of reaction system
PACS:
S 682.1+1
DOI:
-
Abstract:
In order to obtain the optimal SSR-PCR system for Chrysanthemum moriforlium，an orthogonal diagram L16(45) experimental design was employed to evaluate five factors (template DNA，Mg2+，dNTP，primer and Taq DNA polymerase) at four different levels.The fully random single factor experiment was used to select the optimal level for each factor to optimize the SSR-PCR amplification system.The results showed that an optimal SSR-PCR system for chrysanthemum was obtained as following：60 ng DNA template，2.0 mmol/L Mg2+，0.1 mmol/L dNTP，0.3 μmol/L primer，1 U Taq DNA polymerase in 25 μL reaction system.The optimal annealing temperature for SSR-PCR reaction system was determined as 53.1℃ by gradient PCR.The suitable thermal cycling conditions with initial melting at 95℃ for 5 min，followed by 35 cycles at 94℃ for 50 s，53.1℃ for 50 s，72℃ for 50 s；then keep the reaction mixture at 4℃ after a final extension step of 72℃ for 8 min.The optimized system would be effective as a solid foundation for chrysanthemum SSR analysis．

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Last Update: 2014-09-16