Bpu14 I is an enzyme from Bacillus pumilus 14 with a recognition sequence of TTCGAAAAGCTT. One unit of the enzyme can hydrolyze 1 ?g of Lambda DNA in a 50 ?l reaction at 37°C in 1 hour. It is stable in a storage buffer with Tris-HCl, NaCl, EDTA, 2-mercaptoethanol, and glycerol. After overdigestion, 95% of DNA fragments can be ligated. The enzyme has no nonspecific activity after prolonged incubation with DNA. It can be inactivated at 65°C for 20 minutes. The concentration is 10000 u.a./ml, and the working buffer is G with Tris-HCl, MgCl2, NaCl, and DTT.
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