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Isolation of Cosmids using Eppendorf's Perfectprep Plasmid ,,, Mini Kit

Isolation of Cosmids using Eppendorf's Perfectprep Plasmid Mini Kit

James Griffith and Kim Vollmer
James Griffith, University of Georgia, Department of Genetics, Athens, GA
Kimberly Vollmer, Brinkmann Instruments, Biosytems Application Lab, Westbury, NY Abstract

Cosmids are commonly used vectors in the lab, providing the capability to hold large fragments of DNA. They often help when creating genomic libraries where small plasmids and large constructs such as BAC's are not appropriate. Purification of cosmids is a necessary step during their use; Eppendorf's Perfectprep Plasmid Mini Kit is a rapid and effective way of purifying cosmid DNA from bacterial culture.

Introduction

Cosmids have become important tools for molecular biologists, filling a void left between small plasmid vectors and large vector constructs such as BAC's. Large vectors such as cosmids are extremely useful in creating genomic libraries with a capacity to hold fragments of DNA ranging from 30kb and 45kb DNA [1]. Cosmid structures were created from a hybrid of both phage and plasmid traits [2]. The intrinsic characteristics of cosmids allow them to undergo efficient ligation, transfection, replication and selection; providing the exclusivity needed for creating genomic libraries. The characterization of DNA inserts ligated into cosmids is one of the most common downstream applications when creating a library. Due to the size of cosmids it is important to find a method that will purify the cosmid without shearing or excessive loss of DNA. Eppendorf's Perfectprep Plasmid Mini Kit is very versatile, providing an
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