Reaction & Storage Conditions Reaction Conditions: 1X NEBuffer 4 Supplemented with 80 μM S-adenosylmethionine (SAM) and 100 μg/ml Bovine Serum Albumin Incubate at
65°C.
1X NEBuffer 4: 20 mM Tris-acetate 50 mM potassium acetate 10 mM Magnesium Acetate 1 mM Dithiothreitol
pH 7.9 @ 25°C
Unit Definition: One unit is defined as the amount of enzyme required to protect 1 μg λ DNA in 1 hour at 65°C in a total reaction volume of 20 μl against cleavage by TaqI restriction endonuclease.
Protection Assay Conditions:TaqI Methyltransferase is incubated with 1 µg λ DNA in 20 µl 1X NEBuffer 4, 100 µg/ml BSA and 80 µM S-adenosylmethionine, for 1 hour at 65°C. The extent of protection by TaqI Methyltransferase is determined by the addition of 30 µl 1X NEBuffer 4 and 10 units of TaqI restriction endonuclease. Incubation for 1 hour at 65°C is followed by analysis on an agarose gel.
Concentration: 10,000 units/ml
Storage Conditions: 10 mM Tris-HCl 100 mM NaCl 1 mM Dithiothreitol 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol
pH 7.4 @ 25°C
Storage Temperature: -20°C
Notes Usage notes:
TaqI Methyltransferase gives 25% activity at 37°C.
Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product.
16-Hour Incubation:: Incubation of 600 units of TaqI Methyltransferase with 1 μg of BstEII-digested λ DNA in 50 μl NEBuffer 2 at 65°C resulted in no detectable endonuclease contamination.
Exonuclease Activity:: Incubation of 600 units of TaqI Methyltransferase with 1 μg sonicated 3H DNA (105 cpm/μg) for 4 hours at 65°C in 50 μl NEBuffer 2 released < 0.1% of the total radioactivity.
