An Efficient Method for Flanking Sequence Isolation in Barley

被引：7

作者：

Brown, Ryan H. ^{[1
]}

Dahleen, Lynn ^{[2
]}

Bregitzer, Phil ^{[1
]}

机构：

[1] ARS, USDA, Natl Small Grains Germplasm Res Facil, Aberdeen, ID 83210 USA

[2] ARS, USDA, No Crop Sci Lab, Fargo, ND 58102 USA

来源：

CROP SCIENCE | 2012年 / 52卷 / 03期

关键词：

GENOME; DNA; PCR;

D O I：

10.2135/cropsci2011.10.0560

中图分类号：

S3 [农学（农艺学）];

学科分类号：

0901 ;

摘要：

A polymerase chain reaction (PCR)-based adapter ligation method was developed to determine native barley (Hordeum vulgare L.) sequences flanking Dissociation (Ds) insertions and barley expressed sequence tags (ESTs). This method is simple and efficient, with the majority (similar to 70%) of queries returning valid sequence information. The success of this method can be traced to experimental protocols that control sample cross contamination and that segregate specific products from nonspecific products. This report describes the protocol in detail, quantifies its efficiency to enable comparisons to future modifications, and discusses specific problems that may be addressed to enable further improvements to this method.

引用

页码：1229 / 1234

页数：6

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