Suárez Viana Manrique, Clarke David Higginson, López-Canovas Lilia, Riverón Ana María
Molecular Biology Department, Cuban Neurosciences Center, Havana, Cuba.
Prep Biochem Biotechnol. 2008;38(1):40-50. doi: 10.1080/10826060701774338.
DNA molecules suitable for amplification by Polymerase Chain Reaction were obtained by immobilizing whole blood or isolated leukocytes and incubating the immobilized cells for one hour with the known non-enzymatic solution described for preparing intact DNA molecules for PFGE. Cell immobilization was done in agarose gels and punches of 1.2 mm of diameter had the amount of DNA needed for amplifying chromosomal and mitochondrial sequences, many times. The approach was successfully used in preparing DNA molecules from multiple samples in flat-bottom 96-well ELISA plates. The procedure is simple and does not demand special conditions for sample transportation or conservation; thus, it should be useful to collect and process samples under field conditions in epidemiological studies.
通过固定全血或分离的白细胞，并将固定化细胞与用于制备用于脉冲场凝胶电泳（PFGE）的完整DNA分子的已知非酶溶液孵育一小时，获得了适合通过聚合酶链反应进行扩增的DNA分子。细胞固定在琼脂糖凝胶中，直径1.2毫米的打孔块含有多次扩增染色体和线粒体序列所需的DNA量。该方法成功用于从平底96孔酶联免疫吸附测定（ELISA）板中的多个样品制备DNA分子。该程序简单，不需要特殊的样品运输或保存条件；因此，它应该有助于在流行病学研究的现场条件下收集和处理样品。
