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báo cáo khoa học: " Atomic Force Microscope nanolithography on chromosomes to generate single-cell genetic probes"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Atomic Force Microscope nanolithography on chromosomes to generate single-cell genetic probes | Di Bucchianico et al. Journal of Nanobiotechnology 2011 9 27 http www.jnanobiotechnology.eom content 9 1 27 JOURNAL OF NANOBIOTECHNOLOGY RESEARCH Open Access Atomic Force Microscope nanolithography on chromosomes to generate single-cell genetic probes 1 1 1 1 2 2 Sebastiano Di Bucchianico Anna M Poma Maria F Giardi Luana Di Leandro Francesco Valle Fabio Biscarini and Dario Botti 1 Abstract Background Chromosomal dissection provides a direct advance for isolating DNA from cytogenetically recognizable region to generate genetic probes for fluorescence in situ hybridization a technique that became very common in cyto and molecular genetics research and diagnostics. Several reports describing microdissection methods glass needle or a laser beam to obtain specific probes from metaphase chromosomes are available. Several limitations are imposed by the traditional methods of dissection as the need for a large number of chromosomes for the production of a probe. In addition the conventional methods are not suitable for single chromosome analysis because of the relatively big size of the microneedles. Consequently new dissection techniques are essential for advanced research on chromosomes at the nanoscale level. Results We report the use of Atomic Force Microscope AFM as a tool for nanomanipulation of single chromosomes to generate individual cell specific genetic probes. Besides new methods towards a better nanodissection this work is focused on the combination of molecular and nanomanipulation techniques which enable both nanodissection and amplification of chromosomal and chromatidic DNA. Cross-sectional analysis of the dissected chromosomes reveals 20 nm and 40 nm deep cuts. Isolated single chromosomal regions can be directly amplified and labeled by the Degenerate Oligonucleotide-Primed Polymerase Chain Reaction DOP-PCR and subsequently hybridized to chromosomes and interphasic nuclei. Conclusions Atomic force microscope can be easily used to visualize and to .