Seol Yeonee, Neuman Keir C
Laboratory of Single Molecule Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 50, Room 3517, 50 South Drive, Bethesda, 20892, MD, USA.
Methods Mol Biol. 2018;1665:297-316. doi: 10.1007/978-1-4939-7271-5_16.
Magnetic tweezers is a versatile yet simple single-molecule manipulation technique that has been used to study a broad range of nucleic acids and nucleic acid-based molecular motors. In this chapter, we combine micro-mirror-based total internal reflection microscopy with a magnetic tweezers instrument, permitting simultaneous single-molecule visualization and mechanical manipulation. We provide a simple method to calibrate the evanescent wave penetration depth via supercoiling of DNA with a fluorescent nanodiamond-labeled magnetic bead and a complementary method employing a surface-immobilized fluorescent nanodiamond.
磁镊是一种通用且简单的单分子操纵技术，已被用于研究广泛的核酸和基于核酸的分子马达。在本章中，我们将基于微镜的全内反射显微镜与磁镊仪器相结合，实现了单分子的同步可视化和机械操纵。我们提供了一种简单的方法，通过用荧光纳米金刚石标记的磁珠使DNA超螺旋来校准倏逝波穿透深度，以及一种使用表面固定的荧光纳米金刚石的互补方法。
