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Interactome-Seq: A Protocol for Domainome Library Construction, Validation and Selection by Phage Display and Next Generation Sequencing
Published on: October 3, 2018
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Next-generation sequencing of phage-displayed peptide libraries
Wadim L Matochko1, Ratmir Derda
1Department of Chemistry, Alberta Glycomis Centre, University of Alberta, 11227 Saskatchewan Dr., Edmonton, AB, Canada, T6G 2G2.
Methods in Molecular Biology (Clifton, N.J.)
|January 25, 2015
Summary
Next-generation sequencing (NGS) enhances the selection of phage display ligands by detecting rare clones. This study details a method for sequencing Ph.D.-7 phage libraries using Ion Torrent technology and custom analysis.
Area of Science:
- Biotechnology
- Molecular Biology
- Genomics
Background:
- Genetically encoded peptide libraries are crucial for discovering ligands and functional materials.
- Next-generation sequencing (NGS) offers improved ligand selection by detecting low-abundance clones and quantifying copy number changes.
- Limited literature exists on sequencing phage libraries using NGS.
Purpose of the Study:
- To provide a detailed method for sequencing a Ph.D.-7 phage display library.
- To adapt Ion Torrent sequencing for phage library analysis.
- To outline data analysis using a custom Matlab script.
Main Methods:
- Preparation of phage libraries for sequencing.
- Execution of sequencing on the Ion Torrent platform.
- Analysis of sequencing data with a custom Matlab script.
Main Results:
- Demonstration of a feasible method for sequencing phage display libraries.
- Establishment of a workflow for analyzing NGS data from phage libraries.
- Identification of low-abundant clones and quantification of their abundance.
Conclusions:
- NGS, specifically Ion Torrent, is a viable method for analyzing phage display libraries.
- This method enhances ligand discovery and characterization in phage display.
- The described protocol facilitates efficient and comprehensive phage library sequencing.
