High-performance liquid chromatography for the purification of restriction endonucleases, application to BanII, SacI, and SphI

✍ Scribed by V. Bouriotis; A. Zafeiropoulos; Y.D. Clonis

• Publisher: Elsevier Science
• Year: 1987
• Tongue: English
• Weight: 657 KB
• Volume: 160
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(87)90622-1

✦ Synopsis

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.