Sakmanoglu Aslı, Sayin Zafer, Ucan Uckun Sait, Pinarkara Yasemin, Uslu Ali, Erganis Osman
Department of Microbiology, Faculty of Veterinary Medicine, Selcuk University, Konya, Turkey.
Department of Food Technology, Sarayonu Vocational School, Selcuk University, 42430 Konya, Turkey.
J Microbiol Methods. 2017 Sep;140:70-73. doi: 10.1016/j.mimet.2017.07.003. Epub 2017 Jul 12.
Extraction of DNA from Mycoplasma cultured on agar medium is difficult because the plasticity of these microorganisms enables agar penetration. This eventually causes cell loss during harvesting of colonies from the agar surface. Here, we used the GenElute™ gel extraction kit, which is usually used to purify polymerase chain reaction products, for extracting DNA from Mycoplasma. We compared the DNA extraction efficiency of the GenElute™ gel extraction kit from Mycoplasma cynos cultured in agar medium with four other DNA extraction methods. The results were evaluated based on the purity and amount of DNA obtained from one Mycoplasma colony. Eight strains of Mycoplasma cynos isolated from the broncho-alveolar lavage fluid of dogs were used. The GenElute™ gel extraction protocol was the most efficient among all the methods tested in this study as it yielded the highest amount and the purest quality of DNA (199.3±0.744ng/μl) from a single colony. Among the methods tested, the GenElute™ gel extraction method is the most rapid, sensitive, and simple method for DNA extraction from Mycoplasma. This procedure may also prove useful for extracting DNA from other Mycoplasma species.
从在琼脂培养基上培养的支原体中提取DNA很困难，因为这些微生物的可塑性使琼脂能够渗透进去。这最终会导致在从琼脂表面收获菌落的过程中细胞丢失。在这里，我们使用通常用于纯化聚合酶链反应产物的GenElute™凝胶提取试剂盒从支原体中提取DNA。我们将GenElute™凝胶提取试剂盒从在琼脂培养基中培养的犬支原体中提取DNA的效率与其他四种DNA提取方法进行了比较。根据从一个支原体菌落获得的DNA的纯度和数量对结果进行评估。使用了从犬支气管肺泡灌洗液中分离出的八株犬支原体。在本研究测试的所有方法中，GenElute™凝胶提取方案是最有效的，因为它从单个菌落中获得的DNA量最高且质量最纯（199.3±0.744ng/μl）。在测试的方法中，GenElute™凝胶提取法是从支原体中提取DNA最快、最灵敏且最简单的方法。该方法可能也被证明对从其他支原体物种中提取DNA有用。
