Lan Vo Thi Thuong, Loan Pham Thi Thanh, Duong Pham Anh Thuy, Thanh Le Thi, Ha Ngo Thi, Thuan Ta Bich
Faculty of Biology, Hanoi University of Science, Vietnam National University, 334 Nguyen Trai Street, Thanh Xuan, Hanoi, Vietnam.
J Nucleic Acids. 2012;2012:254630. doi: 10.1155/2012/254630. Epub 2012 Feb 12.
DNA ladder is commonly used to determine the size of DNA fragments by electrophoresis in routine molecular biology laboratories. In this study, we report a new procedure to prepare a DNA ladder that consists of 10 fragments from 100 to 1000 bp. This protocol is a combination of routinely employed methods: cloning, PCR, and partial digestion with restriction enzymes. DNA fragments of 100 bp with unique restriction site at both ends were self-ligated to create a tandem repeat. Once being cloned, the tandem repeat was rapidly amplified by PCR and partially digested by restriction enzymes to produce a ladder containing multimers of the repeated DNA fragments. Our procedure for production of DNA ladder could be simple, time saving, and inexpensive in comparison with current ones widely used in most laboratories.
在常规分子生物学实验室中，DNA梯状条带通常用于通过电泳确定DNA片段的大小。在本研究中，我们报告了一种制备DNA梯状条带的新方法，该条带由10个100至1000 bp的片段组成。该方案是常规使用方法的组合：克隆、聚合酶链反应（PCR）和用限制性内切酶进行部分消化。两端具有独特限制性位点的100 bp DNA片段进行自身连接以产生串联重复序列。一旦克隆，串联重复序列通过PCR快速扩增，并用限制性内切酶进行部分消化，以产生包含重复DNA片段多聚体的梯状条带。与目前大多数实验室广泛使用的方法相比，我们生产DNA梯状条带的方法可能简单、省时且成本低廉。
