Upgrading SELEX Technology by Using Lambda Exonuclease Digestion for Single-Stranded DNA Generation 英文参考文献.docVIP

Molecules 2010, 15, 1-11; doi:10.3390/moleculeOPEN ACCESS molecules ISSN 1420-3049 /journal/molecules Article Upgrading SELEX Technology by Using Lambda Exonuclease Digestion for Single-Stranded DNA Generation Meltem Avci-Adali, Angela Paul, Nadja Wilhelm, Gerhard Ziemer and Hans Peter Wendel * Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Calwerstr. 7/1, Tuebingen 72076, Germany ? Author to whom correspondence should be addressed; E-Mail: hans-peter.wendel@med.uni- tuebingen.de; Tel.: +49-7071-2986605; Fax: +49-7071-295369. Received: 19 November 2009; in revised form: 23 December 2009 / Accepted: 23 December 2009 / Published: 24 December 2009 Abstract: 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. Keywords: aptamers; strand separation; streptavidin-coated beads; lambda exonuclease; single-stranded DNA  Molecules 2010, 15 2 Introduction Single-stra