Keeney Stephen
Molecular Diagnostics Centre, Central Manchester University Hospitals Foundation Trust, Manchester, UK.
Methods Mol Biol. 2011;688:67-74. doi: 10.1007/978-1-60761-947-5_6.
Standard polymerase chain reaction (PCR) protocols amplify relatively small fragments precluding the use of this approach when examining gross rearrangements of DNA. By using combinations of DNA polymerases, which feature either good polymerase activity or error-correction abilities, it is now possible to extend the length of DNA fragment that can be amplified. These "long-PCR" protocols have allowed the development of more rapid and convenient ways to analyse large-scale rearrangements of DNA and in many cases has superseded alternative approaches such as Southern blotting. The protocol described in this chapter illustrates some of the key points to be considered when developing a long PCR protocol.
标准聚合酶链反应（PCR）方案扩增的片段相对较小，因此在检测DNA的大规模重排时无法使用该方法。通过使用兼具良好聚合酶活性或纠错能力的DNA聚合酶组合，现在有可能延长可扩增的DNA片段长度。这些“长片段PCR”方案使得开发更快速、便捷的方法来分析DNA的大规模重排成为可能，并且在许多情况下已取代了诸如Southern印迹等替代方法。本章所述的方案阐述了制定长片段PCR方案时需要考虑的一些要点。
