Minnesota/10 July 2008

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1. Dephosphorylation: Continued with double digestion base vector product from 07-09-2008. Used Antartic phosphoylase to dephosphorylate the base vector. Want to dephosphorylate the base vector to prevent it from closing - want it to stay a linear digested product. After dephosphorylation, the base vector was then incubated @ 37C for 30 minutes. Sample was then placed in a heat bath @ 65C for 15 minutes to inactivate enzyme (phosphotase).
Gene 10x Buffer DNA Phosphotase
Base Vector 5.6uL 50.0uL 1.0uL


NOTE: Total volume in dephosphorylation = 56.6uL