Ok Yeon Jeong, You Hee Sang, Lee Song Hee, Sung Ho Joong, Kang Hee-Gyoo, Hyun Sung Hee
1Department of Biomedical Laboratory Science, Eulji University, 77, Gyeryong-ro, 771 beon-gil, Jung-gu, Daejeon, 34824 Republic of Korea.
2Department of Senior Healthcare, Graduate School, BK21 Plus Program, Eulji University, 77, Gyeryong-ro, 771 beon-gil, Jung-gu, Daejeon, 34824 Republic of Korea.
Indian J Microbiol. 2020 Jun;60(2):206-213. doi: 10.1007/s12088-019-00854-6. Epub 2020 Jan 7.
For bacteria sampling studies, various collection methods have been used to identify bacteria. To obtain accurate information about bacteria, high quality samples should be obtained. In order to obtain a high quality sample, a stable and large number of DNA copies must be collected. This study compared the efficiency of different methods of bacterial gDNA extraction and bacteria collection according to swabbing solution volumes and types. The efficiency of bacterial genomic DNA extraction was compared using a AccuPrep Genomic DNA Extraction kit, a QIAamp DNA Mini kit, and a MOBIO DNeasy PowerSoil kit. The DNA Mini kit was shown to extract the highest amount of gDNA, and sub-experiments were conducted using this kit. Phosphate-buffered saline and phosphate-buffered saline with 0.1% Tween 20 were used as collection solutions of various volumes (0, 40, 50, 60, 70, 80, 90, 100, 110, and 120 μL) using cotton swabs. Bacteria collection efficiency was highest when 70 μL PBS was used. The target strains collected in this experiment were and , and these were quantified using the number of colony-forming units, DNA concentrations, and the number of DNA copies. These results can be used to efficiently bacterial collection for experiments in various fields.
在细菌采样研究中,已使用多种采集方法来鉴定细菌。为了获得有关细菌的准确信息,应获取高质量的样本。为了获得高质量的样本,必须采集稳定且大量的DNA拷贝。本研究根据拭子溶液的体积和类型,比较了不同细菌基因组DNA提取方法和细菌采集方法的效率。使用AccuPrep基因组DNA提取试剂盒、QIAamp DNA Mini试剂盒和MOBIO DNeasy PowerSoil试剂盒比较了细菌基因组DNA提取的效率。结果表明,DNA Mini试剂盒提取的基因组DNA量最高,并使用该试剂盒进行了子实验。使用棉签将磷酸盐缓冲盐水和含0.1%吐温20的磷酸盐缓冲盐水用作不同体积(0、40、50、60、70、80、90、100、110和120μL)的采集溶液。使用70μL PBS时细菌采集效率最高。本实验采集的目标菌株为 和 ,并使用菌落形成单位数量、DNA浓度和DNA拷贝数对这些菌株进行了定量。这些结果可用于在各个领域的实验中高效地进行细菌采集。