Hylis Miroslav, Weiser Jaroslav, Oborník Miroslav, Vávra Jirí
Department of Parasitology and Laboratory of Electron Microscopy, Faculty of Science, Charles University, Prague, Czech Republic.
J Invertebr Pathol. 2005 Mar;88(3):257-60. doi: 10.1016/j.jip.2005.02.004. Epub 2005 Mar 23.
DNA from 19 species of microsporidia was isolated and amplified from infected host tissue that were originally prepared between the years 1946 and 1996. The smears, on glass microscope slides, were either Giemsa-stained or unstained. Methanol-fixed, Giemsa-stained smears proved to be suitable for DNA isolation; DNA was amplified from only two of 14 unstained slides. The isolated DNA was successfully amplified in PCRs using small subunit and large subunit rDNA primers and sequenced. The high efficiency of DNA isolation demonstrates the usefulness of archival and type collection slides for some molecular biology and molecular taxonomy studies.
从1946年至1996年间制备的受感染宿主组织中分离并扩增了19种微孢子虫的DNA。玻璃显微镜载玻片上的涂片，有的经过吉姆萨染色，有的未染色。甲醇固定、吉姆萨染色的涂片被证明适合用于DNA分离；14张未染色的载玻片中只有两张成功扩增出了DNA。使用小亚基和大亚基rDNA引物，通过聚合酶链式反应（PCR）成功扩增了分离出的DNA并进行了测序。DNA分离的高效率证明了存档玻片和模式标本玻片在一些分子生物学和分子分类学研究中的实用性。
