July 3rd, 2016
•We present a protocol for the functional assessment of comprehensive single-site saturation mutagenesis libraries of proteins utilizing high-throughput sequencing. Importantly, this approach uses orthogonal primer pairs to multiplex library construction and sequencing. Representative results using TEM-1 β-lactamase selected at a clinically relevant dosage of ampicillin are provided.
Related Videos
Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells
A High Throughput Screen for Biomining Cellulase Activity from Metagenomic Libraries
Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
Competitive Genomic Screens of Barcoded Yeast Libraries
Use of a Robot for High-throughput Crystallization of Membrane Proteins in Lipidic Mesophases
Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans
High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
Generation of High Quality Chromatin Immunoprecipitation DNA Template for High-throughput Sequencing (ChIP-seq)
High Throughput Microinjections of Sea Urchin Zygotes
Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing