DNA sensor's selectivity enhancement and protection from contaminating nucleases due to a hydrated ionic liquid

被引：9

作者：

Tateishi-Karimata, Hisae ^{[1
]}

Pramanik, Smritimoy ^{[1
]}

Sugimoto, Naoki ^{[1
,2
]}

机构：

[1] Konan Univ, Frontier Inst Biomol Engn Res, Kobe, Hyogo 6500047, Japan

[2] Konan Univ, Grad Sch Frontiers Innovat Res Sci & Technol FIRS, Kobe, Hyogo 6500047, Japan

来源：

ANALYST | 2015年 / 140卷 / 13期

关键词：

T BASE-PAIRS; CHEMICAL-STABILITY; PRIMER ELONGATION; G-QUADRUPLEX; TRIPLEX; MISMATCHES; MOTIF; PH; THERMODYNAMICS; FLUORESCENCE;

D O I：

10.1039/c5an00545k

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The thermodynamic stability of certain mismatched base pairs has made the development of DNA sequence sensing systems challenging. Thus, the stability of fully matched and mismatched DNA oligo-nucleotides in the hydrated ionic liquid choline dihydrogen phosphate (choline dhp) was investigated. Mismatched base pairs were significantly destabilized in choline dhp relative to those in aqueous buffer. A molecular beacon that forms a triplex with a conserved HIV-1 sequence was then designed and tested in choline dhp. The molecular beacon specifically detected the target duplex via triplex formation at concentrations as low as 1 pmol per 10 mu L with 10000-fold sequence selectivity. Moreover, the molecular beacon was protected from a contaminating nuclease in choline dhp, and DNAs in aqueous solutions were not sufficiently stable for practical use.

引用

页码：4393 / 4398

页数：6