Negri R, Costanzo G, Di Mauro E
Centro Acidi Nucleici (CNR), Rome, Italy.
Anal Biochem. 1991 Sep 2;197(2):389-95. doi: 10.1016/0003-2697(91)90409-m.
A chemical method for the determination of DNA sequence is presented. Heating of DNA, labeled at a single 3' extremity, in the presence of formamide results in efficient cleavage of phosphodiester bonds 3' of A, G, and C residues. The relative efficiency is A = G greater than C. The bias between A and G is solved by a simple pretreatment (photoreaction in methylene blue) followed by heating in formamide. The entire procedure does not require any intermediate purification step or handling of hazardous chemicals and allows determination of the sequence on only two electrophoretic lanes. Its simplicity and rapidity favor automatization.
本文介绍了一种用于测定DNA序列的化学方法。在甲酰胺存在的情况下，加热单链3'端标记的DNA会导致A、G和C残基3'端的磷酸二酯键有效断裂。相对效率为A = G大于C。通过简单的预处理（亚甲蓝光反应）然后在甲酰胺中加热，可解决A和G之间的偏差。整个过程不需要任何中间纯化步骤或处理危险化学品，并且仅在两条电泳泳道上即可测定序列。其简单性和快速性有利于自动化。
