Small-Scale Isolation of Genomic DNA from Streptomyces Mycelia or Spores

Author:

Kutchma Alecksandr J.1,Roberts Mark A.1,Knaebel David B.1,Crawford Don L.1

Affiliation:

1. University of Idaho, Moscow, ID, USA

Abstract

We have developed a small-scale method for isolating high-quality chromosomal DNA from Streptomyces species. The entire procedure may be carried out in 2-mL microcentrifuge tubes in one day. It has been tested both quantitatively and qualitatively to ensure reliability and reproducibility. DNA yields from a variety of Streptomyces species ranged from 30 μg DNA/50 mg cells to over 225 μg DNA/50 mg cells. We used the method to isolate DNA from cells grown in liquid culture, on solid media and from spore suspensions. DNA yields and quality were assessed by spectrophotometry, restriction endonuclease digestion and random-amplified polymorphic DNA-PCR (RAPD-PCR) analysis. These confirmed that this procedure is an efficient method for isolating large amounts of highquality DNA from a wide range of Streptomyces species.

Publisher

Future Science Ltd

Subject

General Biochemistry, Genetics and Molecular Biology,Biotechnology

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