关键词: Actinobacteria Antimicrobial analyses DNA amplification technique DNA extraction method DNA–DNA hybridization Whole genome sequencing

来  源:   DOI:10.18502/ijm.v14i2.9187   PDF(Pubmed)

Abstract:
UNASSIGNED: DNA extraction is an important step of any molecular experiment. DNA could not be easily extracted from members of actinomycetes by the usual methods of lysis. Due to the low efficiency of the conventional DNA extraction methods, development of an effective technique for DNA extraction of actinobacteria in emergency cases seems to be necessary. Since most of the DNA extraction techniques and commercial kits are not efficient enough to extract DNA from actinobacteria, this study was conducted to improve an efficient method obtained from conventional one to extract DNA from this group of bacteria.
UNASSIGNED: DNA extraction was performed using five methods (an improved method, Invisorb Spin Plant Mini Kit, EZ-10 Spin Column, Sarrbrucken method (HZI, Germany) and Kirby Bauer\'s method). To evaluate the quantity and quality of extracted genomic DNA, UV absorbance of all samples and efficiency of polymerase chain reaction (PCR) were evaluated.
UNASSIGNED: Overall, the results revealed that the highest quantity (up to 4000 ng/μl) and good quality of DNA was obtained using introduced DNA extraction method.
UNASSIGNED: Results indicated that recently introduced improved method was more efficient for extraction of DNA from actinobacteria for DDH (DNA-DNA hybridization) test and for those require the high concentration of DNA.
摘要:
DNA提取是任何分子实验的重要步骤。通过通常的裂解方法不能容易地从放线菌成员中提取DNA。由于传统的DNA提取方法效率低,在紧急情况下,开发一种有效的放线菌DNA提取技术似乎是必要的。由于大多数DNA提取技术和商业试剂盒不足以从放线菌中提取DNA,进行这项研究是为了改进从常规细菌中提取DNA的有效方法。
使用五种方法进行DNA提取(一种改进的方法,Invisorb旋转植物迷你套件,EZ-10旋转柱,Sarrbrucken方法(HZI,德国)和柯比·鲍尔的方法)。为了评估提取的基因组DNA的数量和质量,评估所有样品的UV吸光度和聚合酶链反应(PCR)的效率。
总的来说,结果表明,使用引入的DNA提取方法可以获得最高的DNA数量(高达4000ng/μl)和良好的质量。
结果表明,最近引入的改进方法对于从放线菌中提取DNA进行DDH(DNA-DNA杂交)测试以及需要高浓度DNA的方法更有效。
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