Directed Evolution Library Creation: Methods and Protocols 

1-58829-285-1

978-1-58829-285-8

F. Arnold and G. Georgiou, ed., Humana Press, 2003, 232 pp., hard cover

Part I. Mutagenesis and Recombination Methods
1. Generating Mutant Librarites Using Error-Prone PCR
2. Preparing Libraries in Escherichia coli
3. Preparing Libraries in Saccharomyces cerevisiae
4. Creating Random Mutagenesis Librareis by Megaprimer PCR of Whole Plasmid (MEGAWHOP)
5. Construction of Designed Protein Libraries Using Gene Assembly Mutagenesis
6. Production of Randomly Mutated Plasmid Libraries Using Mutator Strains
7. Evolution of Microorganisms Using Mutator Plasmids
8. Random Insertion and Deletion Mutagenesis
9. Random Oligonucleotide Mutagenesis
10. Saturation Mutagenesis
11. DNA Shuffling
12. Family Shuffling with Single-Stranded DNA
13. In Vitro DAN Recombination by Random Priming
14. Staggered Extention Process (StEP) IN Vitro Recombination
15. RACHITT: Gene Family Shuffling by Random Chimeragenesis on Transient Templates
16. The Creation of ITCHY Hybrid Protein Libraries
17. Preparation of SCRATCHY Hybrid Protein Libraries: Size- and In-Frame Selection of Nucleic Acid Sequences
18. Sequence Homology-Independent Protein Recombination (SHIPREC)
19. Producing Chimeric Genes by CLERY: In Vitro and In Vivo Recombination
PART II. Analysis of Library Diversity
20. Analysis of Shuffled Libraries by Oligonucleotide Probe Hybridization
21. Sequence Mapping of Combinatorial Libraries on Macro- or Microarrays: Experimental Design of DNA Arrays
22. Sequence Mapping of Combinatorial Libraries on Macro- or Microarrays: Bioinformatic Treatment of Data