|Table of Contents|

Establishment of ISSRPCR Reaction System of Cornus walteri(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2013年22期
Page:
114-119
Research Field:
Publishing date:

Info

Title:
Establishment of ISSRPCR Reaction System of Cornus walteri
Author(s):
LI Na; ZHANG Cun-xu; LIU Wei; SONG Li-wei; WANG Hao; YANG Ling
College of Forestry,Northwest Agricultural and Forestry University,Yangling,Shaanxi 712100
Keywords:
Cornus walteri; ISSR; single factor with two circular tests; system optimization; primer screening
PACS:
S 792. 99
DOI:
-
Abstract:
Using leaves of Cornus walteri as material,ISSR reaction conditions of Cornus walteri were optimized systematically via single factor with two circular tests,and the optimal ISSR reaction system and thermal cycling conditions of Cornus walteri were established.The results showed that the optimal ISSR reaction system included a total volume 25 μL containing 2.5 μL 10×Buffer,50 ng template DNA,1.5 U Taq DNA polymerase,2.0 mmol/L MgCl2,0.6 μmol/L primer,0.15 mmol/L dNTPs.Thermal cycling conditions were as follows:denaturation 5 min at 94℃;38 cycles of 50 s at 94℃,60 s at 48.8～59.1℃,1.5 min at 72℃;and a final extension of 10 min at 72℃ at the end of the amplification.Based on the optimized reaction system,17 primers with superior stability and polymorphism were screened out from 100 ISSR primer candidates.Subsequently,annealing temperatures of selected primers were subjected to further optimization.As a consequence,the optimized system in this study may provide technological base and reference for identification of germplasm resources,genetic diversity and molecular breeding of Cornus walter.

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Last Update: 2014-08-07