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Establishment and Optimization of the Method of ISSR Fingerprinting Marker on Limonium Sinense(PDF)

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2012年03期
Page:
93-97
Research Field:
生命科学
Publishing date:

Info

Title:
Establishment and Optimization of the Method of ISSR Fingerprinting Marker on Limonium Sinense
Author(s):
Ding Ge1; Zhang Daizhen2; Yu Yanqiu2; Zhao Lingling1; Zhang Beibei1
1.Department of Chemical and Biological Engineering,Yancheng Institute of Technology,Yancheng 224003,China
Keywords:
Limonium sinense; ISSR; reaction system; orthogonal optimization
PACS:
S567.239;R282.7
DOI:
-
Abstract:
To obtain the optimal amplification of ISSR conditions for Limonium sinense，the effects of the concentrations of DNA templates，magesium，dNTP，Taq DNA polymerase and annealing tempreture were tested and optimized by orthogonal designed experiment． The reliable ISSR － PCR systems for L． sinense was established by analyzing the reasons for occurrence of differential bands and optimizing reaction conditions． The optimal amplification conditions for L． sinense was 25μL PCR reaction volume，10 × Buffer was 2. 5 μL，the concentration of magnesium was 2 mmol /L，dNTP was 100 μmol /L，the concentration of DNA templates was 20 ng，Taq DNA polymerase was 1． 5 U，primer was 0. 4 μmol /L． The appropriate annealing temperature was 52℃ ～ 58℃． The ISSR-PCR systems，which were established in this paper for studying L． sinense，could provide clear reliable abundant polymorphisms molecular markers and were proved suitable for studying genetic diversity and conservation of L． sinense．

References:

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Last Update: 2013-03-11