Application of High Resolution Melting Analysis in Soybean Ecotilling(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:

2011年05期

Page:

843-846,850

Research Field:

Publishing date:

Info

Title:

Application of High Resolution Melting Analysis in Soybean Ecotilling

Author(s):

JIAO Li¹; LIU Xiang-guo²; YANG Chun-ming²; WANG Hu-yi³; HAO Dong-yun²; 3; WANG Qing-yu¹

1.College of Plant Science,Jilin University,Changchun 130062;

2.Biotechnology Center,Jilin Academy of Agricultural Sciences,Changchun 130124;?

3.College of Life Science,Jilin University,Changchun 130012,Jilin,China

Keywords:

Soybean; High resolution melting; Ecotilling; SNP

PACS:

S565.1

DOI:

10.11861/j.issn.1000-9841.2011.05.0843

Abstract:

Identification of allelic variants in natural populations by Ecotilling has became a rapid Single nucleotide polymorphisms(SNP) discovery method,but it is not convenient for detection.High resolution melting(HRM)analysis is a rapid,sensitive and high-throughput method that can be used in the examination of SNP.In this paper,HRM techniques was used in Ecotilling to detect the SNP of IFS1 exons in 256 soybean cultivars.The results of HRM was verified by direct sequencing.The effect of homogenization and non-homogenization of the template on the testing was discussed and the result showed that the former had more remarkable effect.The testing effect of different proportions of mixed samples was compared and found that 1∶1 and 1∶4 mixed samples were optimal.The comparison between the testing results of exon regions of IFS1 by HRM and results of PCR products sequencing showed the accuracy of fifth and seventh sections of exons were 91.2% and 100%,respectively.It demonstrates that the soybean-cultivating Ecotilling established by the use of HRM is a new method for SNP detection with high sensitivity, throughput and low cost.

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