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  • Title: Pseudo-affinity column chromatography based rapid purification procedure for T7 RNA polymerase.
    Author: Das M, Dasgupta D.
    Journal: Prep Biochem Biotechnol; 1998 Nov; 28(4):339-48. PubMed ID: 9805352.
    Abstract:
    Based on the observation that T7 RNAP binds reversibly to the polyaromatic sulphonated triazine dye, cibacron blue, we have developed a rapid purification procedure for T7 RNAP. It employs chromatography of the ammonium sulfate fraction through a blue sepharose column, which has the dye coupled to the solid sepharose support. The enzyme can be eluted by 2M NaCl or 1M NaCl together with 1 mM UTP. These methods are compared with another purification procedure using ion-exchange column chromatography. All of them yield essentially pure T7 RNAP with high specific activity.
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