A simplified quick microbial genomic DNA extraction via freeze-thawing cycles.
Mol Biol Rep
; 47(1): 703-709, 2020 Jan.
Article
en En
| MEDLINE
| ID: mdl-31713008
ABSTRACT
Effective isolation of high-quality genomic DNA is one of the essential steps in molecular biology, biochemistry, and genetic studies. Here we describe a simplified procedure based on repeated freeze-thawing cycles to isolate genomic DNA from different organisms of microbes (Burkholderia pyrrocinia JK-SH007, Bacillus pumilus HRl0, Botrytis cinerea) and nematodes (Bursaphelenchus xylophilus). The DNA extraction buffer includes 10% of CTAB; 4% of NaCl (W/V); 20 mM of ethylenediamine tetraacetic acid; 100 mM of Tris-HCl, pH 8.0 and 1% of polyvinylpyrrolidone. The released DNA was purified from the mixture using a phenol/chloroform mixture and precipitated in 70% ethanol to remove proteins, carbohydrates, phenols, RNA, etc. Our method is a reproducible, simple, and rapid technique for routine DNA extractions from various microorganisms and nematodes. Furthermore, the low cost of this method could be an economic benefit to large-scale studies.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
ADN Bacteriano
/
Técnicas Microbiológicas
/
Fraccionamiento Químico
Límite:
Animals
Idioma:
En
Año:
2020
Tipo del documento:
Article