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BiochemistrybioRxiv
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An Efficient and Cost-effective Purification Methodology for Cas9 Nuclease

A. C. T. Teng, M. Tavassoli, S. Shrestha, K. Lindsay, E. Ivakine, R. Cohn, J. P. Santerre, A. O. Gramolini

Abstract

With an ever-increasing demand for laboratory-grade Cas9 proteins by many groups advancing the use of CRISPR technology, a more efficient and scalable process for generating the proteins, coupled with rapid purification methods is in urgent demand. Here, we introduce a modified methodology for rapid purification of active SaCas9 protein within 24 hours. The product has over 90% protein purity. The simplicity and cost-effectiveness of such methodology will enable general labs to produce a sizable amount of Cas9 proteins, further accelerating the advancement of CRISPR/Cas9-based research.

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