Open AccessEffects of the turn-back primer on intermediate product generation in isothermal DNA amplification
Author(s) -
Yuki Tanaka,
Yoshinobu Kimura,
Yuichiro Mitani,
Yosuke Kawai,
Alexander Lezhava,
Shohei Noma,
Michihira Tagami,
Jun Kawai,
Yoshihide Hayashizaki,
Kengo Usui
Publication year - 2010
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000113566
Subject(s) - primer (cosmetics) , loop mediated isothermal amplification , turn (biochemistry) , dna , rolling circle replication , biology , chemistry , microbiology and biotechnology , genetics , dna polymerase , biochemistry , organic chemistry
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.