Mte I

Supplied in:

10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0,1 mM EDTA; 7 mM  2-mercaptoethanol; 200 µg/ml BSA; 50% glycerol.

Reaction Conditions:

1XSEBuffer W. 

Incubate at  55°C.

1XSEBuffer W  (pH 8.5 @ 25ºC)

10 mM Tris-HCl,100  mM NaCl, 10 mM MgCl2,1 mM DTT

Unit Definition:

One unit is defined as the amount of enzyme required to hydrolyze completely 1 μg of linearized plasmid pHspAI10/DriI+M.Fsp4HI in 1 hour at 55°C in a total reaction volume of 50μl. pHspAI10/DriI+M.Fsp4HI is a plasmid pHspAI10, which is linearized with DriI, and, additionally, modified with Fsp4HI DNA methyltransferase pHspAI10 carries a gene of HspAI DNA methyltransferase, that modifies the sequence . 5`-GCGC-3`, producing 5`-G(5mC)GC-3`. M.Fsp4HI modifies the sequence 5`-GCNGC-3`, producing 5`-G(5mC)NGC-3`. A substrate pHspAI10/DriI+M.Fsp4HI includes one site

5`G(5mC)G(5mC)NG(5mC)G(5mC)-3`/3`(5mC)G(5mC)GN(5mC)G(5mC)G-5`,which is MteI canonical site [1]. The enzyme activity depends on a number and positions of methylated nucleotides in the recognition sequence. For example, MteI cuts the recognition site with six 5-methylcytosines, but the enzyme activity is reduced for more that one order [1]

Quality Control Assays

16-Hour Incubation:

No detectable degradation of 1μg of Lambda DNA was observed after incubation with 10 units of enzyme for 16 hours at 55°C in a total reaction volume of 50 μl.

Oligonucleotide Assay:

No detectable degradation of a single- and double-stranded oligonucleotide was observed after incubation with 10 units of enzyme for 3 hours.

Enzyme properties

Activity in SEBuffers:

SEBuffer B            25- 50%

SEBuffer G           75-100%

SEBuffer O           75-100%

SEBuffer W                100%

SEBuffer Y             50- 75%

SEBuffer ROSE          100%

When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve a complete digestion.

Reagents Supplied with Enzyme:

10XSEBuffer W

Heat Inactivation: 

No