Ambrose B J, Pless R C
Biochemistry. 1985 Oct 22;24(22):6194-200. doi: 10.1021/bi00343a025.
A novel approach to sequence analysis of end-labeled, defined DNA fragments, using a single chemical cleavage procedure and electrophoretic separation in a single lane, has been developed. Prolonged treatment with hot aqueous piperidine results in partial cleavage of the DNA at all positions; the relative propensity for this cleavage is different for the various bases in the DNA. The hydrolysate is resolved on a DNA sequencing gel, and the distribution of radioactivity in the electrophoretic lane is analyzed (a) in terms of differential peak heights of the radioactive bands and (b) in terms of the spacings between successive bands. Simultaneous application of these two base-characteristic criteria allows the deduction of the nucleotide sequence with an accuracy approaching that of the established four-lane methods of DNA sequencing.
一种用于末端标记的特定DNA片段序列分析的新方法已经开发出来，该方法使用单一化学切割程序，并在单泳道中进行电泳分离。用热的哌啶水溶液长时间处理会导致DNA在所有位置部分切割；这种切割的相对倾向因DNA中的各种碱基而异。水解产物在DNA测序凝胶上进行分离，并分析电泳泳道中放射性的分布：(a) 根据放射性条带的差异峰高，以及(b) 根据连续条带之间的间距。同时应用这两个碱基特征标准，可以推导出核苷酸序列，其准确性接近已确立的四泳道DNA测序方法。
