Vizard D, Hadman M, Bianca D, Manzer D
International Biotechnologies, New Haven, CT 06511.
Biotechniques. 1990 Apr;8(4):430-7.
A simplified method of DNA sequencing by dideoxy chain termination is developed that approaches a single-step protocol. Utilizing the sequencing advantages contributed by a thermophilic polymerase and a guanine analog, stable sequencing reaction concentrates have been obtained that readily perform the entire sequencing reaction simply by adding prepared DNA to each of the four reaction concentrates required by this method. The mechanics and dynamics of these reactions have been investigated and the capacity of these reactions to withstand normal user variation is demonstrated. This study focuses on one form of this simplified method embodied in the FASTaq DNA sequencing kit.
开发了一种通过双脱氧链终止进行DNA测序的简化方法，该方法接近单步操作流程。利用嗜热聚合酶和鸟嘌呤类似物带来的测序优势，已获得稳定的测序反应浓缩物，只需将制备好的DNA添加到该方法所需的四种反应浓缩物中，即可轻松完成整个测序反应。对这些反应的机制和动力学进行了研究，并证明了这些反应承受正常用户操作差异的能力。本研究聚焦于FASTaq DNA测序试剂盒中所体现的这种简化方法的一种形式。
