Affiliations 

  • 1 Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Penang, Malaysia
Anal Biochem, 2012 Dec 1;431(1):54-6.
PMID: 22975202 DOI: 10.1016/j.ab.2012.08.025

Abstract

The quality of a nucleotide-based library such as a synthetic antibody library is highly dependent on the diversity available. Diversity can be generated using degenerate oligonucleotides introduced during gene assembly. Conventional approaches to gene assembly are not efficient for oligonucleotides with long stretches of degeneracy. We propose an efficient alternative for simultaneous introduction of three randomized regions in a synthetic antibody gene via temperature cascading. The strategy takes advantage of DNA reannealing kinetics. The strategy can be adopted for generating diversity of gene inserts during the construction of nucleotide-based libraries.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.