Rapid cloning of genes and promoters for functional analyses

Schenk, Peer M. (2014). Rapid cloning of genes and promoters for functional analyses. In Agnelo Furtado and Robert J. Henry (Ed.), Cereal Genomics: Methods and Protocols (pp. 123-132) New York, United States: Humana Press. doi:10.1007/978-1-62703-715-0_11


Author Schenk, Peer M.
Title of chapter Rapid cloning of genes and promoters for functional analyses
Title of book Cereal Genomics: Methods and Protocols
Place of Publication New York, United States
Publisher Humana Press
Publication Year 2014
Sub-type Critical review of research, literature review, critical commentary
DOI 10.1007/978-1-62703-715-0_11
Open Access Status
Year available 2014
Series Methods in Molecular Biology
ISBN 9781627037143
9781627037150
ISSN 1064-3745
1940-6029
Editor Agnelo Furtado
Robert J. Henry
Volume number 1099
Chapter number 11
Start page 123
End page 132
Total pages 10
Total chapters 24
Collection year 2015
Language eng
Abstract/Summary Next-generation sequencing has resulted in a massive fl ow of new information predicting the existence of many new genes, their putative promoters, as well as long and small noncoding RNA. However, this is currently largely unmatched by functional studies. A cost-effective and high-throughput cloning system for PCR products and synthetic sequences was therefore developed to allow the rapid evaluation of coding and noncoding sequences in functional expression and reporter assays. Unlike traditional cloning approaches that involve subcloning or a special recipient vector and special fl anking sequences, this protocol describes a rapid and cost-effective method for the direct insertion into the vector of choice. Restriction enzymes are only needed once to prepare the vector, which is blunt ended and dephosphorylated, and can then serve as the recipient vector for many hundreds of sequences to be tested. Examples are provided of how this method can be used to rapidly reveal functionality of regulatory genes, promoters, and microRNAs.
Keyword Functional assays
Functional genomics
Gene mining
Library construction
Q-Index Code BX
Q-Index Status Confirmed Code
Institutional Status UQ

 
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Created: Tue, 25 Mar 2014, 20:39:00 EST by Prof Peer Schenk on behalf of Qld Alliance for Agriculture and Food Innovation