将DNA聚合酶I用作RNA依赖性DNA聚合酶的条件。
Conditions for using DNA polymerase I as an RNA-dependent DNA polymerase.
作者信息
Gulati S C, Kacian D L, Spiegelman S
出版信息
Proc Natl Acad Sci U S A. 1974 Apr;71(4):1035-9. doi: 10.1073/pnas.71.4.1035.
Abstract
Conditions are described for using Escherichia coli DNA polymerase I for synthesizing complementary DNA copies of natural RNA molecules, which are suitable for use in hybridization experiments. The molar ratio of enzyme to template is critical; below a certain level, synthesis is not observed. Hybrids formed with the complementary DNA are of comparable specificity and stability to those formed with complementary DNAs synthesized by viral RNA-directed DNA polymerase. Synthesis of dA-dT polymers, a common occurrence with this enzyme, can be eliminated by including distamycin in the reaction mixture.
摘要
描述了使用大肠杆菌DNA聚合酶I合成天然RNA分子互补DNA拷贝的条件，这些拷贝适用于杂交实验。酶与模板的摩尔比至关重要；低于一定水平时，无法观察到合成。与互补DNA形成的杂交体在特异性和稳定性方面与由病毒RNA指导的DNA聚合酶合成的互补DNA形成的杂交体相当。通过在反应混合物中加入偏端霉素，可以消除该酶常见的dA-dT聚合物的合成。
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