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

Alecksandr Kutchma | Zendy; Mark A. Roberts | Zendy; David B. Knaebel | Zendy; Don L. Crawford | Zendy

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

Author(s) -

Alecksandr Kutchma,

Mark A. Roberts,

David B. Knaebel,

Don L. Crawford

Publication year - 1998

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/98243st05

Subject(s) - biology , dna , streptomyces , restriction enzyme , rapd , genomic dna , spore , polymerase chain reaction , mycelium , dna extraction , streptomycetaceae , endonuclease , genomic library , microbiology and biotechnology , actinomycetales , chromatography , genetics , bacteria , gene , chemistry , botany , base sequence , population , demography , sociology , genetic diversity

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 micrograms DNA/50 mg cells to over 225 micrograms 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 high-quality DNA from a wide range of Streptomyces species.

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