Optimization of ISSR-PCR System and Selection of Primers for Horticultural Varieties of Clematis(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Title:: Optimization of ISSR-PCR System and Selection of Primers for Horticultural Varieties of Clematis

Author(s):: WANG Nan; WANG Jin; LI Zongyan; YANG Kunmei; HE Hui; XIAO Zhendong; （College of Landscape Architecture,Southwest Forestry University，Kunming，Yunnan 650224)

Keywords:: Clematis cultivation; ISSR-PCR reaction system; primer screening

Abstract:: A Clematis garden species （Clematis ‘Gravetype Beauty’）was chosen as the experiment material to built the genetic map based on the ISSR molecule marker technology.The ‘Gravetye Beauty’ genomic DNA was used as the DNA model,and simple factor design of experiment was used to study the impacts of model NDA concentration,primer concentration,and Master Mix on ISSR-PCR reaction.The results showed that the optimum system was DNA model 5 ng,primer concentration 0.8 μmol/L,Master Mix 12 μL and the whole reaction system was 25 μL.Using this system,11 primers with clear stripe and good polymorphism were selected from 38 ISSR primers.

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Last Update: 2016-03-02