纸表微生物的简易模型与DNA提取方法评价

A Simple Model and DNA Extraction of Microbes from Paper Surface

微生物是造成古籍善本和纸质档案损伤的主要原因之一,近年来不依赖于实验室培养的高通量测序成为研究纸表微生物的常用方法.但是纸表微生物的含量往往偏低,DNA提取常依赖于昂贵的进口试剂盒,成本过高.本文首先构建了一个标准的纸表微生物体系,用传统棉签擦拭法获取该纸表微生物样品,再利用改良的化学法、机械破壁法、酶消解法3种方式提取样品中微生物总DNA并比较其浓度与纯度,最后利用PCR扩增验证提取效果.结果表明,酶消解法提取所得浓度最高,化学法次之,机械法最低,但DNA纯度正好相反.经PCR验证,化学法的扩增效果较好.综合上述指标,改良的化学法更适宜提取纸表微生物样品中的总DNA.

Microorganism is the current research hotspot as one of the important reasons caused serious damage for ancient books and documents.Since the amount of microbes on the paper surface is limited,their genome DNA has to be extracted with the expensive kits.In order to develop a cheaper and easier method,we first constructed a simple system to simulate the microbiota niche on the paper surface.The samples were collected with the cotton tip and the genome DNA were extracted with three different methods.The concentration and purity of DNA were compared with each other and the quality of DNA was evaluated with PCR amplification.The results showed that the concentration of the DNA extracted with enzyme method was the highest than those of chemical method and physical method,but the purity was just on the opposite.PCR results indicated that the DNA quality with the chemical method was better than other two methods,which implied that the chemical method was with potential to be adopted in the DNA extraction from paper surface.