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Primer-Free Aptamer Selection mit einem zufälligen DNA-Bibliothek
1Department of Pathology, Hershey Medical Center,Pennsylvania State University, 2Department of Chemistry,Pennsylvania State University, 3Departments of Pathology, and Biochemistry and Molecular Biology, Hershey Medical Center,Pennsylvania State University, 4Materials Research Institute,Pennsylvania State University
Kapitel
- 00:04Titel
- 00:12Introduction
- 01:39Preparation of PF Library-DNA Fragments
- 03:26Preparation of S100B-bound Beads
- 04:55Selection & Recovery of Aptamers
- 06:40TOPO Cloning and Sequencing
- 07:17Sandwich Binding Assays
- 09:11Representative Results
- 10:35Conclusion
SELEX-Protokolle umfassen mehrere Runden der Selektion, von denen jeder Regeneration von Liganden, die wiederum erfordern feste Primersequenzen flankierenden die zufällige Bibliothek Regionen erfordern. Diese festen Primersequenzen können mit der Auswahl (false positive und negative) stören. Hier präsentieren wir eine Grundierung-free-Protokoll.
Tags
Primer-Free Aptamer SelectionRandom DNA LibraryAptamersOligonucleotidesDNARNAAffinitiesAntibodiesIn Vitro Selection ProcessSystematic Evolution Of Ligands By EXponential Enrichment (SELEX)TargetsSmall MoleculesProteinsMacromoleculesIn Vivo Diagnostic ApplicationsTherapeutic ApplicationsSpecificitySynthesizedProcessing ConditionsImmunogenicitySizeNucleotides (nt)Nucleic Acid LibrariesRandomized RegionsPrimer SitesSelection Process ComplicationsBioinformatics Approaches
