Jensen M A, Zagursky R J, Trainor G L, Cocuzza A J, Lee A, Chen E Y
Central Research and Development Department, E.I. du Pont de Nemours & Company (Inc.), Wilmington, DE 19898.
DNA Seq. 1991;1(4):233-9. doi: 10.3109/10425179109020778.
Significant improvements in the quality of DNA sequencing data have been shown when deoxyadenosine triphosphate (dATP) is replaced by 7-deaza-2'-deoxyadenosine triphosphate (c7dATP). The use of c7dATP in conjunction with 7-deaza-2'-deoxyguanosine triphosphate (c7dGTP) further decreases anomalies in electrophoretic mobility which are caused by compressions involving G and/or A residues. This effect is observed for both isotope-based and fluorescence-based sequencing approaches. Replacing dATP with c7dATP also results in a higher degree of uniformity in the frequency of chain termination reactions, when such terminations involve the incorporation of fluorescence-labeled dideoxynucleotides by T7 polymerase. These improvements in the gel-resolution and distribution of chain-terminated DNA products result in higher accuracy in both manual and automated base assignment.
当用7-脱氮-2'-脱氧三磷酸腺苷（c7dATP）取代脱氧三磷酸腺苷（dATP）时，DNA测序数据的质量有了显著提高。将c7dATP与7-脱氮-2'-脱氧鸟苷三磷酸（c7dGTP）结合使用，进一步减少了由涉及G和/或A残基的压缩引起的电泳迁移异常。在基于同位素和基于荧光的测序方法中均观察到这种效果。当链终止反应涉及T7聚合酶掺入荧光标记的双脱氧核苷酸时，用c7dATP取代dATP还会导致链终止反应频率的更高程度的均匀性。链终止DNA产物在凝胶分辨率和分布方面的这些改进，使得手动和自动碱基分配的准确性更高。
