Nucleic acid library construction using synthetic DNA constructs

Methods Mol Biol. 2005:288:359-78. doi: 10.1385/1-59259-823-4:359.

Abstract

This chapter outlines seven synthetic and molecular biology techniques that allow the controlled synthesis of nucleic acid libraries. Specifically: (1) The high-diversity chemical synthesis of point mutations; (2) the high-diversity chemical synthesis of point deletions; (3) the split-bead approach for constructing point mutation or deletion libraries with limited sequence diversity; (4) pool deprotection, gel purification, and quality-control techniques; (5) large-scale polymerase chain reaction amplification for the generation of high-diversity double-stranded deoxyribonucleic acid libraries; (6) type II restriction enzyme digestion techniques for the construction of long-sequence libraries containing minimal fixed sequence; and (7) extension techniques for the rapid synthesis of long, low-diversity oligonucleotide sequences.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry*
  • Nucleic Acids / chemical synthesis*
  • Nucleic Acids / genetics
  • Point Mutation
  • Polymerase Chain Reaction

Substances

  • Nucleic Acids
  • DNA