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1Department of Health Sciences,Università del Piemonte Orientale & IRCAD, Novara, Italy, 2Institute of Biomedical Technologies,National Research Council, Segrate, Milan, Italy, 3Institute of Biomedical Technologies,National Research Council, Bari, Italy, 4Department of Life Sciences,University of Trieste, Italy, 5Institute of Genetic and Biomedical Research,National Research Council, Rozzano, Milan, Italy, 6Humanitas Clinical and Research Center, Rozzano, Milan, Italy
Chapters
- 00:04Title
- 01:19Construction of the ORF Library
- 02:38Preparation of the Filtering Vector
- 03:40Ligation and Transformation
- 06:47pFILTER-ORF Library Validation and Collection
- 09:10Phage Library Deep Sequencing Platform, Index PCR Followed by Qualitative and Quantitative Evaluation of the Sequencing Library
- 10:07Results: Use of a Phagemid Vector to Construct, Characterize and Select a Domainome Library
- 11:23Conclusion
Les protocoles décrits permettent la construction, la caractérisation et la sélection (contre une cible de choix) d’une bibliothèque « domainome » fabriquée à partir de n’importe quelle source d’ADN. Ceci est réalisé par un pipeline de recherche qui combine différentes technologies : affichage de phage, reporter pliant et séquençage de génération suivant avec un outil web pour l’analyse des données.
