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Small-scale isolation of genomic DNA from Streptomyces mycelia or spores. 1998

A J Kutchma, and M A Roberts, and D B Knaebel, and D L Crawford
University of Idaho, Moscow, USA.

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.

UI MeSH Term Description Entries
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture
D004269 DNA, Bacterial Deoxyribonucleic acid that makes up the genetic material of bacteria. Bacterial DNA
D013053 Spectrophotometry The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.
D013171 Spores, Bacterial Heat and stain resistant, metabolically inactive bodies formed within the vegetative cells of bacteria of the genera Bacillus and Clostridium. Bacterial Spores,Bacterial Spore,Spore, Bacterial
D013302 Streptomyces A genus of bacteria that form a nonfragmented aerial mycelium. Many species have been identified with some being pathogenic. This genus is responsible for producing a majority of the ANTI-BACTERIAL AGENTS of practical value.
D015183 Restriction Mapping Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA. Endonuclease Mapping, Restriction,Enzyme Mapping, Restriction,Site Mapping, Restriction,Analysis, Restriction Enzyme,Enzyme Analysis, Restriction,Restriction Enzyme Analysis,Analyses, Restriction Enzyme,Endonuclease Mappings, Restriction,Enzyme Analyses, Restriction,Enzyme Mappings, Restriction,Mapping, Restriction,Mapping, Restriction Endonuclease,Mapping, Restriction Enzyme,Mapping, Restriction Site,Mappings, Restriction,Mappings, Restriction Endonuclease,Mappings, Restriction Enzyme,Mappings, Restriction Site,Restriction Endonuclease Mapping,Restriction Endonuclease Mappings,Restriction Enzyme Analyses,Restriction Enzyme Mapping,Restriction Enzyme Mappings,Restriction Mappings,Restriction Site Mapping,Restriction Site Mappings,Site Mappings, Restriction
D019105 Random Amplified Polymorphic DNA Technique Technique that utilizes low-stringency polymerase chain reaction (PCR) amplification with single primers of arbitrary sequence to generate strain-specific arrays of anonymous DNA fragments. RAPD technique may be used to determine taxonomic identity, assess kinship relationships, analyze mixed genome samples, and create specific probes. RAPD Technique,RAPD-PCR,Arbitrarily Primed PCR Reaction,RAPD Technic,Random Amplified Polymorphic DNA Technic,RAPD Technics,RAPD Techniques

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