Hin6I, Restriction Enzyme
10mM Tris-HCl (pH 7.4 at 25 degree C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods the Storage Buffer should be used.
Digestion of Agarose-embedded DNA
A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
AciI, Bsp119I, Bsu15I, Hin1I, HpaII, MaeII, MspI, NarI, Psp1406I, TaqI, XmiI
Hin6I does not cut Gm5CGC. Blocked by CG methylation.
Labeled Oligonucleotide (LO) Assay
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours.
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Hin6I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 2.0uM. More than 95% of these can be recut.
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Hin6I.
Stability During Prolonged Incubation
A minimum of 0.1 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37 degree C.
10mM potassium phosphate (pH 7.4 at 25 degree C), 100mM NaCl, 1mM EDTA, 7mM 2-mercaptoethanol, 0.2mg/ml BSA and 50% glycerol.
Enzyme is inactivated by incubation at 65 degree C for 20min.
One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37 degree C in 50ul of assay buffer.