The enzyme cleaves only C5-methylated DNA and doesn't cut unmodified DNA.
10 mM Tris-HCl (pH 7.5); 100mM NaCl; 0.1mM EDTA; 7mM 2-mercaptoethanol; 50% glycerol; Store at -20°C
1X SEBuffer Bis I
Incubate at 37°C
Warranty period for the enzyme storage at -20˚C is two years from the date of the last assay indicated on the enzyme vial.
1X SEBuffer BisI (pH 9.0 @ 25ºC)
10 mM Tris-HCl, 150 mM KCl, 10 mM MgCl2, 1 mM DTT
One unit is defined as the amount of enzyme required to cleave 1 pmol of the double-stranded oligonucleotide with the following
5' GCTTGTACTTTAG(5mC)GGCATTGATTCTCACCACG 3'
3' CGAACATGAAATCGC(5mC)GTAACTAAGAGTGGTGC 5'
in 1 hour at 37°C in a total reaction volume of 20μl.
Reagents supplied with enzyme:
10X SEBuffer Bis I
Yes (65°C for 20 minutes)
Quality Control Assays
No nonspecific activity was detected after incubation of 1 μg of pFsp4HI1 DNA (BamHI
digest) with 1 unit of BisI for 16 hours at 37°C. The pFsp4HI1 plasmid carries a gene for Fsp4HI DNA-methyltransferase, which modifies DNA
No detectable degradation of a single- and double-stranded oligonucleotide was
observed after incubation with 1 units of enzyme for 3 hours.
Activity in SEBuffers:
SEBuffer B 10-25%
SEBuffer G 25-50%
SEBuffer O 50-75%
SEBuffer W 75-100%
SEBuffer Y 50-75%
SEBuffer ROSE 30%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
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