Upgrading SELEX Technology by Using Lambda Exonuclease Digestion for Single-Stranded DNA Generation
| Autor: | Angela Paul, Meltem Avci-Adali, Gerhard Ziemer, Hans Peter Wendel, Nadja Wilhelm |
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| Rok vydání: | 2009 |
| Předmět: |
Time Factors
Aptamer Blotting Western aptamers strand separation DNA Single-Stranded Pharmaceutical Science Lambda exonuclease single-stranded DNA Biology Nucleic Acid Denaturation Article DNA sequencing Analytical Chemistry lcsh:QD241-441 Magnetics streptavidin-coated beads lambda exonuclease chemistry.chemical_compound lcsh:Organic chemistry Drug Discovery Sodium Hydroxide Biotinylation Denaturation (biochemistry) Phosphorylation Physical and Theoretical Chemistry SELEX Aptamer Technique Organic Chemistry Single-strand conformation polymorphism Molecular biology Microspheres Exodeoxyribonucleases chemistry Biochemistry Chemistry (miscellaneous) Molecular Medicine Streptavidin DNA microarray DNA Systematic evolution of ligands by exponential enrichment |
| Zdroj: | Molecules; Volume 15; Issue 1; Pages: 1-11 Molecules, Vol 15, Iss 1, Pp 1-11 (2009) Molecules |
| ISSN: | 1420-3049 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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