Highly Parallel Magnetic Tweezers by Targeted DNA Tethering
Highly Parallel Magnetic Tweezers by Targeted DNA Tethering
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DOI：
10.1021/nl203299e
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发表时间：
2011-12-01
期刊：
影响因子：
10.8
通讯作者：
Dekker, Cees
中科院分区：
文献类型：
--
作者：
De Vlaminck, Iwijn;Henighan, Thomas;Dekker, Cees
单分子力谱方法，如磁钳和光钳，已经成为详细研究DNA-酶相互作用的生物力学方面的强大工具。由于在单个实验中通常只处理单个DNA分子，这些方法存在数据吞吐量低的问题。在这里，我们报告了一种新的方法，通过微接触打印DNA末端结合标签来定向、非随机地固定规则阵列中的DNA系留磁珠。我们发现，由于DNA珠链排列成规则的阵列而导致密度的增加，可以引起磁镊子实验中数据吞吐量的一个数量级的提高。我们证明了这项技术在镊子实验中的适用性，在这些实验中，多达450个珠子被同时跟踪，从一次实验运行中产生了357个分子的DNA力学统计数据。我们的技术为千分子力谱实验铺平了道路，使研究DNA蛋白质相互作用中的罕见事件成为可能，并从单个实验运行中获取大型统计数据集。
Single-molecule force-spectroscopy methods such as magnetic and optical tweezers have emerged as powerful tools for the detailed study of biomechanical aspects of DNA-enzyme interactions. As typically only a single molecule of DNA is addressed in an individual experiment, these methods suffer from a low data throughput. Here, we report a novel method for targeted, nonrandom immobilization of DNA-tethered magnetic beads in regular arrays through microcontact printing of DNA end-binding labels. We show that the increase in density due to the arrangement of DNA-bead tethers in regular arrays can give rise to a one-order-of-magnitude improvement in data-throughput in magnetic tweezers experiments. We demonstrate the applicability of this technique in tweezers experiments where up to 450 beads are simultaneously tracked in parallel, yielding statistical data on the mechanics of DNA for 357 molecules from a single experimental run. Our technique paves the way for kilo-molecule force spectroscopy experiments, enabling the study of rare events in DNA protein interactions and the acquisition of large statistical data sets from individual experimental runs.