Optimization of ISSR Amplification Conditions in Ancient Camphor-trees(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:

2013年07期

Page:

116-120

Research Field:

Publishing date:

Info

Title:

Optimization of ISSR Amplification Conditions in Ancient Camphor-trees

Author(s):

HUANG Meng¹; 2; CHEN Shang-xing¹; 2; YANG Guang-yao¹; 2; WANG Zong-de¹; 2; JI Chun-feng¹; 2

1.College of Landscape and Art，Jiangxi Agricultural University，Nanchang，Jiangxi 330045；

2.Jiangxi Provincial Key Laboratory for Bamboo Germplasm Resources and Applications，Jiangxi Agricultural University，Nanchang，Jiangxi 330045

Keywords:

ancient camphor-trees; ISSR; reaction condition; optimization

PACS:

S 792.23

DOI:

-

Abstract:

The genomic DNA of ancient camphor-trees was extracted from leaves by improved CTAB method.With the design of single factor experiment method，the influences of seven factors including Mg²⁺，dNTP，DNA content in the template，Taq DNA polymerase，amounts of primer，deionized formamide，and annealing temperature on PCR amplification were analyzed.The results showed that the best ISSR-PCR reaction system of ancient camphor-trees was as follows：50 ng DNA template，2.5 mmol/L MgCl₂，0.20 mmol/L dNTPS，1.0 U Taq polymerase，0.5 μmol/L primer and 2% formamide deionized in 20 μL reaction volume.The results would lay a technology foundation for further study of the genetic diversity of ancient camphor-trees with ISSR molecular marking.

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