Accelerated isothermal nucleic acid amplification in betaine-free reaction

被引：25

作者：

Ma, Cuiping ^{[1
]}

Wang, Yifan ^{[1
]}

Zhang, Pansong ^{[1
]}

Shi, Chao ^{[2
]}

机构：

[1] Qingdao Univ Sci & Technol, Coll Marine Sci & Biol Engn, Key Lab Sensor Anal Tumor Marker, Minist Educ, Qingdao 266042, Peoples R China

[2] Qingdao Univ, Coll Life Sci, Qingdao 266071, Peoples R China

来源：

ANALYTICAL BIOCHEMISTRY | 2017年 / 530卷

基金：

中国国家自然科学基金;

关键词：

Betaine; Isothermal amplification; Molecular barrier; Associate rate constant; POCT; OF-CARE DIAGNOSTICS; VIRUS; DNA; SYSTEM;

D O I：

10.1016/j.ab.2017.04.017

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Betaine was used as a common additive to isothermal nucleic acid amplification reactions because of lowering the melting temperature (Tm) of DNA. Herein, we reported a novel finding that betaine was inhibiting the reaction efficiency of isothermal amplification reactions. In this work, we have verified this finding by classical loop-mediated isothermal amplification that the addition of 0.8 M betaine inhibited the efficiency of reaction dropping to approximately 1%. Additionally, we clarified the mechanism of betaine hindering isothermal amplification reactions with a molecular barrier to lower associate rate constant K1 for intermolecular hybridization. This finding would be very significant for studies on the interaction between small molecule substance and DNA, and the development of point-of-care testing because of simplifying reaction system and increasing reaction efficiency. (C) 2017 Elsevier Inc. All rights reserved.

引用

页码：1 / 4

页数：4