Ostrofet Eugeniu, Papini Flávia S, Dulin David
Junior Research Group 2, Interdisciplinary Center for Clinical Research, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Cauerstraße 3, 91058 Erlangen, Germany.
Data Brief. 2020 Mar 12;30:105397. doi: 10.1016/j.dib.2020.105397. eCollection 2020 Jun.
Gene expression is achieved by enzymes as RNA polymerases that translocate along nucleic acids with steps as small as a single base pair, i.e., 0.34 nm for DNA. Deciphering the complex biochemical pathway that describes the activity of such enzymes requires an exquisite spatiotemporal resolution. Magnetic tweezers are a powerful single molecule force spectroscopy technique that uses a camera-based detection to enable the simultaneous observation of hundreds of nucleic acid tethered magnetic beads at a constant force with subnanometer resolution [1,2]. High spatiotemporal resolution magnetic tweezers have recently been reported [3-5]. We present data acquired using a bespoke magnetic tweezers instrument that is able to perform either in high throughput or at high resolution. The data reports on the best achievable resolution for surface-attached polystyrene beads and DNA tethered magnetic beads, and highlights the influence of mechanical stability for such assay. We also present data where we are able to detect 0.3 nm steps along the z-axis using DNA tethered magnetic beads. Because the data presented here are in agreement with the best resolution obtained with magnetic tweezers, they provide a useful benchmark comparison for setup adjustment and optimization.
基因表达是由诸如RNA聚合酶之类的酶来实现的，这些酶沿着核酸移动，移动步长小至单个碱基对，即对于DNA来说为0.34纳米。解读描述此类酶活性的复杂生化途径需要精确的时空分辨率。磁镊是一种强大的单分子力谱技术，它使用基于相机的检测方法，能够在恒定力作用下以亚纳米分辨率同时观察数百个核酸连接的磁珠[1,2]。最近已有关于高时空分辨率磁镊的报道[3 - 5]。我们展示了使用定制磁镊仪器获取的数据，该仪器能够进行高通量或高分辨率操作。这些数据报告了表面附着的聚苯乙烯珠和DNA连接的磁珠可达到的最佳分辨率，并突出了机械稳定性对此类检测的影响。我们还展示了使用DNA连接的磁珠沿z轴检测到0.3纳米步长的数据。由于此处呈现的数据与磁镊获得的最佳分辨率一致，它们为设置调整和优化提供了有用的基准比较。
