An improved method to identify the T-DNA insertion site in transgenic Arabidopsis thaliana genome

An improved method to identify the T-DNA insertion site in transgenic Arabidopsis thaliana genome An improved method to identify the T-DNA insertion site in transgenic Arabidopsis thaliana (Columbia ecotype) genome was presented. Firstly, the pre-adaptor was amplified by PCR from the plasmid pLASC11.12.8 and digested by HindIII to produce the adaptor. After treated with calf intestine alkaline phosphatase, the adaptor was ligated to the genomic restriction digested fragment with the same restriction endonucleases. Then two rounds of PCR (nested-PCR) were carried out and an unknown sequence between the T-DNA and the adaptor was amplified. Further analysis would reveal the accurate site of T-DNA insertion into transgenic A. thaliana genome. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Plant Physiology Springer Journals

An improved method to identify the T-DNA insertion site in transgenic Arabidopsis thaliana genome

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Publisher
Nauka/Interperiodica
Copyright
Copyright © 2007 by Pleiades Publishing, Ltd.
Subject
Life Sciences; Plant Physiology; Plant Sciences
ISSN
1021-4437
eISSN
1608-3407
D.O.I.
10.1134/S1021443707060155
Publisher site
See Article on Publisher Site

Abstract

An improved method to identify the T-DNA insertion site in transgenic Arabidopsis thaliana (Columbia ecotype) genome was presented. Firstly, the pre-adaptor was amplified by PCR from the plasmid pLASC11.12.8 and digested by HindIII to produce the adaptor. After treated with calf intestine alkaline phosphatase, the adaptor was ligated to the genomic restriction digested fragment with the same restriction endonucleases. Then two rounds of PCR (nested-PCR) were carried out and an unknown sequence between the T-DNA and the adaptor was amplified. Further analysis would reveal the accurate site of T-DNA insertion into transgenic A. thaliana genome.

Journal

Russian Journal of Plant PhysiologySpringer Journals

Published: Nov 7, 2007

References

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