期刊论文

Xiong, Xing-Yao

英文

DNA AND CELL BIOLOGY

1044-5498

2012

31

4

600-606

10.1089/dna.2011.1379

National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31071826]; Special Research Fund for Doctor's Degree Dissertation in Chinese Universities [20104320120008]; China Postdoctoral Science FoundationChina Postdoctoral Science Foundation [20100471215, 201104473]; Hunan College Foundation [09C497, SCX1033]

本校为第一且通讯机构

We describe a new application of megaprimer polymerase chain reaction (PCR) for constructing a tandemly repeated DNA sequence using the drought responsive element (DRE) from Arabidopsis thaliana as an example. The key feature in the procedure was PCR primers with partial complementarity but differing melting temperatures (T m). The reverse primer had a higher T m, a 3′ end complementary to the DRE sequence and a 5′ region complementary to the forward primer. The initial cycles of the PCR were conducted at a lower primer annealing temperature ...