Kutchma A J, Roberts M A, Knaebel D B, Crawford D L
University of Idaho, Moscow, USA.
Biotechniques. 1998 Mar;24(3):452-6. doi: 10.2144/98243st05.
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.
我们开发了一种从链霉菌属物种中分离高质量染色体DNA的小规模方法。整个过程可以在2毫升微量离心管中于一天内完成。该方法已经过定量和定性测试，以确保可靠性和可重复性。多种链霉菌属物种的DNA产量范围为30微克DNA/50毫克细胞至超过225微克DNA/50毫克细胞。我们使用该方法从液体培养、固体培养基上生长的细胞以及孢子悬浮液中分离DNA。通过分光光度法、限制性内切酶消化和随机扩增多态性DNA-PCR（RAPD-PCR）分析对DNA产量和质量进行了评估。这些结果证实，该方法是从多种链霉菌属物种中分离大量高质量DNA的有效方法。
