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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
Chapters
- 00:04Title
- 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 protocollen bestaan uit meerdere ronden van selectie, die elk vereisen regeneratie van gebonden liganden, die op hun beurt behoefte aan vaste primer sequenties aan weerszijden van de willekeurige bibliotheek regio's. Deze vaste primer sequenties kunnen interfereren met de selectieprocedure (vals positieven en negatieven). Hier presenteren wij een primer-vrij protocol.
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
