DNA Cloning and Sequencing

Abstract

The plasmid DNA is cleaved with an enzyme and joined in vitro to foreign DNA; the resulting recombinant plasmids are then used to transform bacteria. The plasmid vectors must be carefully chosen and processed to minimize the effort required to identify and characterize recombinants. This chapter provides guidelines for preparation of DNA fragment for cloning, transformation into chemically competent host, and selection of positive clones. The write-up will also describe basic methods used in the cloning of PCR amplified rRNA gene into appropriate vector and followed by sequencing.