Efficient Method for Flanking Sequence Isolation in Barley

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 (-70%) of queries returning...

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Bibliographic Details
Published inCrop science Vol. 52; no. 3; pp. 1229 - 1234
Main Authors Brown, Ryan H, Dahleen, Lynn, Bregitzer, Phil
Format Journal Article
LanguageEnglish
Published Madison, WI Crop Science Society of America 01.05.2012
The Crop Science Society of America, Inc
Subjects
Agronomy. Soil science and plant productions
barley
Biological and medical sciences
cross contamination
expressed sequence tags
Fundamental and applied biological sciences. Psychology
Hordeum vulgare
polymerase chain reaction
Barley
Monocotyledones
Hordeum vulgare
Gramineae
Angiospermae
Spermatophyta
Flanking sequence
Cereal crop
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Summary: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 (-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.
Bibliography:http://dx.doi.org/10.2135/cropsci2011.10.0560
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ISSN:1435-0653
0011-183X
1435-0653
DOI:10.2135/cropsci2011.10.0560