Bouriotis V, Zafeiropoulos A, Clonis Y D
Anal Biochem. 1987 Jan;160(1):127-34. doi: 10.1016/0003-2697(87)90622-1.
Conventional fractionation methods are time consuming, thus they prolong the time required to process low-stability restriction enzymes. We now report a rapid and effective two-step chromatographic method that affords high purity endonucleases in a short time. Accordingly, an inexpensive chromatographic adsorbent such as phosphocellulose or dyed agarose in the first step is coupled to a high-performance ion exchanger, namely, MonoQ, in the second step. The purification schemes reported here are now in routine use to prepare high-purity BanII, SacI, and SphI as judged by the "overdigestion" and "cut-ligate-recut" stringent quality tests.
传统的分级分离方法耗时较长，因此延长了处理低稳定性限制酶所需的时间。我们现在报告一种快速有效的两步色谱法，该方法可在短时间内提供高纯度的核酸内切酶。相应地，第一步使用廉价的色谱吸附剂，如磷酸纤维素或染色琼脂糖，第二步与高性能离子交换剂，即MonoQ相结合。根据“过度消化”和“切割-连接-再切割”严格质量测试判断，此处报道的纯化方案目前已常规用于制备高纯度的BanII、SacI和SphI。
