Sarah L. Wood

Document Type

Honors Project

First Advisor

Dr. Stephen Baron

Degree Award Date

Spring 2006


Gene Library, Streptomyces spp. 5A, BamHI restriction digest, Lucigen's BigEasy Linear Cloning Kit


Bacteriology | Biology | Microbiology


A method for fractionating genomic DNA from Streptomyces spp. 5A DNA into 6.6 to 23 kilobase (kb) fragments was established using a BamHI restriction digest. Construction of a gene library with BamHI digested Streptomyces spp. 5A DNA was attempted using Lucigen®'s BigEasy™ Linear Cloning Kit. However, only one in ten transformants tested proved to be true recombinants while the remaining nine were non-recombinants containing religated vector. The high background of non-recombinants was undesirable. However, the fact that at least one recombinant was found suggests that this cloning method might be optimized to achieve higher proportions of recombinants.