Sachleben R A, Brown G M, Sloop F V, Arlinghaus H F, England M W, Foote R S, Larimer F W, Woychik R P, Thonnard N, Jacobson K B
Chemistry Division, Oak Ridge National Laboratory, TN 37831.
Genet Anal Tech Appl. 1991 Sep;8(6):167-70. doi: 10.1016/1050-3862(91)90057-x.
A method is described for synthesis of a tin reagent, triethylstannylpropanoic acid (TESPA), and its attachment to oligonucleotide primers. Except for the expected mobility retardation, the presence of [116Sn]-TESPA did not affect the sequencing ladder on electrophoresis gels. By using [120Sn]-TESPA and [35S]-dTTP simultaneously in the Sanger procedure, DNA bands on an electrophoresis gel were first located by autoradiography and then by resonance ionization spectroscopy to demonstrate the coincidence of the signals. Previous results using stable isotopes as labels on model compounds are now confirmed by their use in actual DNA sequencing products.
描述了一种合成锡试剂三乙基锡基丙酸（TESPA）及其与寡核苷酸引物连接的方法。除了预期的迁移率降低外，[116Sn]-TESPA的存在并不影响电泳凝胶上的测序梯带。通过在桑格测序法中同时使用[120Sn]-TESPA和[35S]-dTTP，电泳凝胶上的DNA条带首先通过放射自显影定位，然后通过共振电离光谱法定位，以证明信号的重合。以前将稳定同位素用作模型化合物标记的结果现在通过在实际DNA测序产物中的应用得到了证实。
