TY - JOUR
T1 - High yield preparation of genomic DNA from Streptomyces
AU - Nikodinovic, Jasmina
AU - Barrow, Kevin D.
AU - Chuck, Jo-Anne
PY - 2003
Y1 - 2003
N2 - Streptomyces species produce important drugs such as antibiotics, immunosuppressants, and antitumor compounds.The isolation of genomic DNA is imperative for the understanding of the biosynthesis of these compounds and has led to the rational design of new analogs (1−5). Streptomyces are Gram-positive bacteria, making DNA isolation difficult due to their resistance to cell lysis (6,7). Most methods use lysozyme and sodium docecyl sulfate (SDS) for cell disruption. To further increase lysis, glycine is often incorporated into media to minimize peptidoglycan cross-linking; muramidases such as mutanolysin or grinding of mycelia are also commonly used (1,8−10). Compared with DNA isolation methods for Escherichia coli, most methods are time-consuming or low yielding, or give low-quality DNA (1). This report details an improved method for DNA isolation from Streptomyces species using achromopeptidase, lysozyme, and SDS for cell lysis that results in higher yield compared with current standard methods.
AB - Streptomyces species produce important drugs such as antibiotics, immunosuppressants, and antitumor compounds.The isolation of genomic DNA is imperative for the understanding of the biosynthesis of these compounds and has led to the rational design of new analogs (1−5). Streptomyces are Gram-positive bacteria, making DNA isolation difficult due to their resistance to cell lysis (6,7). Most methods use lysozyme and sodium docecyl sulfate (SDS) for cell disruption. To further increase lysis, glycine is often incorporated into media to minimize peptidoglycan cross-linking; muramidases such as mutanolysin or grinding of mycelia are also commonly used (1,8−10). Compared with DNA isolation methods for Escherichia coli, most methods are time-consuming or low yielding, or give low-quality DNA (1). This report details an improved method for DNA isolation from Streptomyces species using achromopeptidase, lysozyme, and SDS for cell lysis that results in higher yield compared with current standard methods.
KW - DNA
KW - achromopeptidase
KW - cell lysis
KW - genomics
KW - lysozyme
KW - streptomyces
UR - http://handle.uws.edu.au:8081/1959.7/45662
UR - http://www.biotechniques.com/biotechniques/multimedia/archive/00010/03355bm05_10134a.pdf
M3 - Article
SN - 0736-6205
JO - Biotechniques: the International Journal of Life Science Methods
JF - Biotechniques: the International Journal of Life Science Methods
ER -