An improved method to identify the T-DNA insertion site in transgenic Arabidopsis thaliana genome |
| |
Authors: | S. C. Huang H. Z. Liu G. H. He F. G. Yu |
| |
Affiliation: | (1) College of Life Science, Nanjing University, Hankou Road 22, Nanjing, 210093, China |
| |
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. This text was submitted by the authors in English. |
| |
Keywords: | Arabidopsis thaliana T-DNA localization adaptor nested-PCR |
本文献已被 SpringerLink 等数据库收录! |