10 mM Bis-Tris Propane-HCl (pH 6.5 at 37°C), 10 mM MgCl2, 100 mM KCl and 0.1 mg/ml BSA.
Incubate at 37°C.
Ligation and Recleavage
After 50-fold overdigestion with Bpu10I, more than 90% of the pBR322 DNA fragments can be ligated in a reaction mixture containing 20-40 u of T4 DNA Ligase/1 µg of fragments and 10% PEG. No more than 50% of these can be recut due to asymmetric recognition sequence of Bpu10I. The remaining uncleaved ligation products may be cut by Eco81I (SauI) and Bpu1102I (EspI).
Storage Buffer
Bpu10I is supplied in:
10 mM Tris-HCl (pH 7.5 at 25°C), 200 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/ml BSA and 50% (v/v) glycerol.
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