RAPID METHOD FOR DETERMINING SEQUENCES IN DNA BY PRIMED SYNTHESIS WITH DNA-POLYMERASE
RAPID METHOD FOR DETERMINING SEQUENCES IN DNA BY PRIMED SYNTHESIS WITH DNA-POLYMERASE
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DOI：
10.1016/0022-2836(75)90213-2
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发表时间：
1975-01-01
影响因子：
5.6
通讯作者：
COULSON, AR
中科院分区：
文献类型：
--
作者：
SANGER, F;COULSON, AR
本文介绍了一种简单、快速的DNA聚合酶引物合成法测定单链DNA核苷酸序列的方法。这取决于在不同限制性核苷三磷酸的条件下使用大肠杆菌DNA聚合酶I和噬菌体T4的DNA聚合酶，并通过丙烯酰胺凝胶上的离子电泳根据大小同时分离产物。以人工合成的十核苷酸A-G-A-A-A-T-A-A-A-A-A-A为引物，用限制性内切酶酶切产物为引物，对噬菌体φ X 174 DNA中的两个序列进行了测定。
A simple and rapid method for determining nucleotide sequences in single-stranded DNA by primed synthesis with DNA polymerase is described. It depends on the use ofEscherichia coliDNA polymerase I and DNA polymerase from bacteriophage T4 under conditions of different limiting nucleoside triphosphates and concurrent fractionation of the products according to size by ionophoresis on acrylamide gels. The method was used to determine two sequences in bacteriophage φX174 DNA using the synthetic decanucleotide A-G-A-A-A-T-A-A-A-A and a restriction enzyme digestion product as primers.