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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
The protocols described allow the construction, characterization and selection (against the target of choice) of a "domainome" library made from any DNA source. This is achieved by a research pipeline that combines different technologies: phage display, a folding reporter and next generation sequencing with a web tool for data analysis.
