McKenna Sean A, Kim Insil, Puglisi Elisabetta Viani, Lindhout Darrin A, Aitken Colin Echeverría, Marshall R Andrew, Puglisi Joseph D
Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.
Nat Protoc. 2007;2(12):3270-7. doi: 10.1038/nprot.2007.480.
RNA synthesis using in vitro transcription by phage T7 RNA polymerase allows preparation of milligram quantities of RNA for biochemical, biophysical and structural investigations. Previous purification approaches relied on gel electrophoretic or gravity-flow chromatography methods. We present here a protocol for the in vitro transcription of RNAs and subsequent purification using fast-performance liquid chromatography. This protocol greatly facilitates production of RNA in a single day from transcription to purification.
利用噬菌体T7 RNA聚合酶进行体外转录来合成RNA，能够制备毫克级的RNA用于生化、生物物理及结构研究。以往的纯化方法依赖于凝胶电泳或重力流色谱法。我们在此介绍一种用于RNA体外转录及随后使用快速高效液相色谱进行纯化的方案。该方案极大地促进了从转录到纯化在一天内完成RNA的制备。
