13] Simultaneous synthesis and biological applications of DNA fragments: An efficient and complete methodology

Abstract

Publisher Summary This chapter discusses the simultaneous synthesis and biological applications of deoxyribo nucleic acid (DNA) fragment that is an efficient and complete methodology. One must first analyze the specific situation and consider factors of confidence, speed, flexibility, budget, available know-how, and annual demand. Suppose the decision was made to synthesize oligonucleotides in-house and to use them for the construction of genes or mutagenesis cassettes, or just for a variety of smaller projects. The basic idea of this approach is rather simple. Instead of using small beads or fibers, as in the classical approaches to peptide and nucleic acid synthesis, this chapter employs “segmental supports,” non interchangeable entities, as the immobilizing support. This type of support allows one to elongate simultaneously those oligonucleotide chains that require the same nucleotide. This chapter concludes with the discussion on the construction of DNA double strands. DNA double strands constructed as described basically represent pieces of DNA built up from small cassettes. New sequence variants are easily made, in high yields, by exchange, insertion, or deletion of two or more fragments. Most of the fragments can be reused many times.