Ronaghi M, Nygren M, Lundeberg J, Nyrén P
Department of Biotechnology, The Royal Institute of Technology (KTH), Stockholm, SE-100 44,
Anal Biochem. 1999 Feb 1;267(1):65-71. doi: 10.1006/abio.1998.2978.
A common problem in conventional DNA sequencing is the occurrence of DNA sequence compressions during gel electrophoresis, leading to misreading of the sequence. These compressions are usually due to secondary structures in the DNA fragment. In this study, we present a non-gel-based DNA sequencing technique that facilitates analysis of such DNA regions. A part of the polymorphic pertussis toxin promoter region in five different Bordetella species was successfully resolved by the new technique. The obtained sequence data revealed four related palindromic sequences. The ability of different DNA polymerases to read through such secondary structures is also described.
传统DNA测序中的一个常见问题是在凝胶电泳过程中出现DNA序列压缩，导致序列误读。这些压缩通常是由于DNA片段中的二级结构造成的。在本研究中，我们提出了一种基于非凝胶的DNA测序技术，有助于分析此类DNA区域。新技术成功解析了五种不同博德特氏菌属物种中多态性百日咳毒素启动子区域的一部分。获得的序列数据揭示了四个相关的回文序列。还描述了不同DNA聚合酶读取此类二级结构的能力。
