Effects of the turn-back primer on intermediate product generation in isothermal DNA amplification. (December 2010)

• Record Type: Journal Article
• Authors: Tanaka, Yuki; Kimura, Yasumasa; Mitani, Yasumasa; Kawai, Yuki; Lezhava, Alexander; Noma, Shohei; Tagami, Michihira; Kawai, Jun; Hayashizaki, Yoshihide; Usui, Kengo
• Is Part Of: Biotechniques. Volume 49:Number 6(2010)
• Page Start: 888
• Page End: 892
• Publication Date: 2010-12
• Subjects: isothermal DNA amplification -- strand displacement -- turn-back primer -- single-molecule amplification -- 454 sequencing; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000113566
• Languages: English
• ISSNs: 0736-6205

Abstract

In DNA amplification, the initial step of copying a target sequence from the template DNA, or the so-called intermediate product generation step, is very important. In examining the turn-back primer (TP)–dependent isothermal DNA amplification (TIA) method, we determined the actual time point of intermediate product generation by extrapolating dsDNA amplification curves. Our results indicate that intermediate product creation is the rate-limiting step in TIA, and good TP design is advantageous for improving the intermediate production process.