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SibEnzyme Ltd. :: Display product information: Bis I
Name
Bis I
Catalog numberE485E486
Package, u.a.40200
Concentration, u.a./ml1000-20001000-2000

Recognition siteG(5mC)^NGC
CGN^(5mC)G
Methylation sensitivityThe enzyme cleaves only C5-methylated DNA.
OriginBacillus subtilis T30
Storage conditions10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20C.
Assayed onDouble-stranded oligonucleotide
5 GCTTGTACTTTA G(5mC)G G C ATTGATTCTCACCACG 3
3' CGAACATGAAAT C G C(5mC)G TAACTAAGAGTGGTGC 5'
Non-specific activityNo nonspecific activity was detected after incubation of 1 μg of pFsp4HI1 DNA (BamHI digest) with 1 unit of BisI for 16 hours at 37C. The pFsp4HI1 plasmid carries a gene for Fsp4HI DNA-methyltransferase, which modifies DNA forming 5'-G(5mC)NGC-3'/3'-CGN(5mC)G-5'.
Optimum temperature37 oC
UnitOne unit is defined as the amount of enzyme required to cleave 1 pmol of the double-stranded oligonucleotide of the above indicated structure in 1 hour at 37C in a total reaction volume of 20 μl.
Inactivation 20 minutes65oC
Optimum SE-bufferSE-buffer BisI (10 mM Tris-HCl (pH 9,0 at 25C); 10 mM MgCl2; 150 mM KCl; 1 mM DTT.)

Enzyme % activity:
BGOWY
10 - 2525 - 5050 - 7575 - 10050 - 75

Note: No detectable degradation of a double-stranded labeled oligonucleotide was observed after incubation with 1 unit of restriction endonuclease BisI for 4 hours at 37C.

References:

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