title: Upgrading SELEX technology by using lambda exonuclease digestion for single-stranded DNA generation.
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.
authors.affiliation: Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Calwerstr. 7/1, Tuebingen 72076, Germany. hans-peter.wendel@med.uni-tuebingen.de
authors.forename: Meltem
authors.initials: M
authors.lastname: Avci-Adali
authors.forename: Angel
authors.initials: A
authors.lastname: Paul
authors.forename: Nadj
authors.initials: N
authors.lastname: Wilhelm
authors.forename: Gerhard
authors.initials: G
authors.lastname: Ziemer
authors.forename: Hans Peter
authors.initials: HP
authors.lastname: Wendel
mesh_terms: D019921:Biotinylation / Q000187:drug effects; D015153:Blotting, Western; D004277:DNA, Single-Stranded / Q000096:biosynthesis*; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D008280:Magnetics; D008863:Microspheres; D009691:Nucleic Acid Denaturation / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D052156:SELEX Aptamer Technique / Q000379:methods*; D012972:Sodium Hydroxide / Q000494:pharmacology; D019809:Streptavidin / Q000378:metabolism; D013997:Time Factors
chemical_list: D004277:DNA, Single-Stranded; D012972:Sodium Hydroxide; D019809:Streptavidin; D005090:Exodeoxyribonucleases