Xu X M, Ikai A
Laboratory of Biodynamics, Tokyo Institute of Technology, Yokohama, Japan.
Biochem Biophys Res Commun. 1998 Jul 30;248(3):744-8. doi: 10.1006/bbrc.1998.9027.
Using a modified tip of the atomic force microscope (AFM), we harvested several strands of genomic DNA from a nanometer region of mouse chromosomes. We have also co-developed a random PCR method to amplify the recovered genomic DNA, in which a single DNA molecule of several kilobasepairs could be amplified efficiently. A subsequent fluorescence in situ hybridization (FISH) indicated that the amplified DNA originally came from the tip-manipulated regions of mouse chromosomes. Several fragments containing unique sequences were identified using Southern hybridization after subcloning the PCR products into pUC18 plasmid. The present results showed a potential application of AFM to genomic analysis.
我们使用原子力显微镜（AFM）的改良探针，从小鼠染色体的纳米区域收获了几条基因组DNA链。我们还共同开发了一种随机PCR方法来扩增回收的基因组DNA，该方法可以高效扩增数千碱基对的单个DNA分子。随后的荧光原位杂交（FISH）表明，扩增的DNA最初来自小鼠染色体的探针操作区域。将PCR产物亚克隆到pUC18质粒中后，通过Southern杂交鉴定出了几个含有独特序列的片段。目前的结果显示了AFM在基因组分析中的潜在应用。
