Upgrading SELEX Technology by Using Lambda Exonuclease Digestion for Single-Stranded DNA Generation

被引：102

作者：

Avci-Adali, Meltem ^{[1
]}

Paul, Angela ^{[1
]}

Wilhelm, Nadja ^{[1
]}

Ziemer, Gerhard ^{[1
]}

Wendel, Hans Peter ^{[1
]}

机构：

[1] Univ Tubingen Hosp, Dept Thorac Cardiac & Vasc Surg, D-72076 Tubingen, Germany

来源：

MOLECULES | 2010年 / 15卷 / 01期

关键词：

aptamers; strand separation; streptavidin-coated beads; lambda exonuclease; single-stranded DNA; POLYMERASE CHAIN-REACTION; PCR AMPLIFICATION; APTAMERS; SELECTION; ENHANCEMENT; TEMPLATES; MOLECULES; SEQUENCE; BETAINE;

D O I：

10.3390/molecules15010001

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The generation of single-stranded DNA (ssDNA) molecules plays a key role in the SELEX (Systematic Evolution of Ligands by EXponential enrichment) combinatorial chemistry process and numerous molecular biology techniques and applications, such as DNA sequencing, single-nucleotide polymorphism (SNP) analysis, DNA chips, DNA single-strand conformation polymorphism (SSCP) analysis and many other techniques. The purity and yield of ssDNA can affect the success of each application. This study compares the two ssDNA production methods, the strand separation by streptavidin-coated magnetic beads and alkaline denaturation and the lambda exonuclease digestion, in regard to the purity of generated ssDNA and the efficiency. Here, we demonstrate the considerable benefits of ssDNA production by lambda exonuclease digestion for in vitro selection of DNA aptamers. We believe that the generation of ssDNA aptamers using this method will greatly improve the success rate of SELEX experiments concerning the recovery of target-specific aptamers.

引用

页码：1 / 11

页数：11

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