Tran-Nguyen Lucy T T, Schneider Bernd
Department of Resources, Plant Industries, Darwin, NT, Australia.
Methods Mol Biol. 2013;938:381-93. doi: 10.1007/978-1-62703-089-2_32.
CsCl-bisbezimide gradient centrifugation is a method to separate phytoplasma from host plant DNA. Bisbenzimide forms a complex with A + T-rich DNA thereby lowering its relative density. During centrifugation the A + T-rich phytoplasma DNA is spatially separated from the less A + T-rich host plant DNA. The difference in buoyant density between phytoplasma DNA and plant DNA varies according to the host-pathogen combination. The phytoplasma DNA forms a distinct band above the host plant DNA and can be collected. Depending on the phytoplasma titer and the scale of extraction, highly purified DNA is obtained in sufficient quantities for the construction of a genomic library, a sequencing project or hybridization studies.
氯化铯-双苯甲酰亚胺梯度离心法是一种从寄主植物DNA中分离植原体的方法。双苯甲酰亚胺与富含A+T的DNA形成复合物，从而降低其相对密度。在离心过程中，富含A+T的植原体DNA在空间上与富含A+T较少的寄主植物DNA分离。植原体DNA和植物DNA之间的浮力密度差异因寄主-病原体组合而异。植原体DNA在寄主植物DNA上方形成一条明显的带，可以收集。根据植原体滴度和提取规模，可获得足够数量的高纯度DNA，用于构建基因组文库、测序项目或杂交研究。
