Ulrich R L, Hughes T A
Department of Microbiology and Molecular Medicine, Clemson University, Clemson, SC 29672, USA.
Lett Appl Microbiol. 2001 Jan;32(1):52-6. doi: 10.1046/j.1472-765x.2001.00866.x.
This study describes a rapid procedure for the isolation of genomic DNA from various Gram-positive bacteria. Species tested included Lactobacillus delbrueckii subsp. lactis ATCC 4797, Lact. acidophilus N2, Staphylococcus aureus, Staph. epidermidis, Propionibacterium jensenii P126, Bacillus pumilus and Enterococcus faecalis. Our technique for chromosomal DNA isolation circumvents the need for phenol-chloroform extractions and caesium chloride gradients. Isolated DNA is consistently greater than 25 kb in size and can be used directly for subcloning, polymerase chain reaction amplification, restriction digestions and library construction.
本研究描述了一种从多种革兰氏阳性菌中快速分离基因组DNA的方法。所测试的菌种包括德氏乳杆菌乳酸亚种ATCC 4797、嗜酸乳杆菌N2、金黄色葡萄球菌、表皮葡萄球菌、詹氏丙酸杆菌P126、短小芽孢杆菌和粪肠球菌。我们用于分离染色体DNA的技术无需进行酚-氯仿抽提和氯化铯梯度离心。分离得到的DNA大小始终大于25 kb，可直接用于亚克隆、聚合酶链反应扩增、限制性酶切消化和文库构建。
