TaqI Methyltransferase

Description

TaqI Methyltransferase modifies the adenine residue (N6) of the sequence TCGA.

Product Source

An E. coli strain that carries the cloned TaqI modification gene from Thermus aquaticus.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
S-adenosylmethionine (SAM)-2032 mM
CutSmart® Buffer-2010X

Properties and Usage

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.

Reaction Conditions

1X CutSmart® Buffer
Supplement with 80 μM S-adenosylmethionine (SAM)
Incubate at 65°C

1X CutSmart® Buffer:
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 μg/ml BSA
pH 7.9 @ 25°C

Protection Assay Conditions:
TaqI Methyltransferase is incubated with 1 µg λ DNA in 20 µl 1X CutSmart Buffer 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 CutSmart Buffer and 10 units of TaqI restriction endonuclease. Incubation for 15 minutes at 65°C is followed by analysis on an agarose gel.

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
100 mM NaCl
1 mM DTT
0.1 mM EDTA
200 μg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Molecular Weight

Theoretical: 47845 daltons

Notes

  1. TaqI Methyltransferase gives 25% activity at 37°C.

References

  1. Hoffman, J.L. (1986). Biochemistry. 25, 4444-4449.

FAQs

  1. Is S-adenosylmethionine (SAM) supplied with the Methyltransferase?
  2. What should be considered if the methylation is not going to completion?
  3. What is the activity of TaqI Methyltransferase at 37°C?
  4. What is the activity of TaqI Methyltransferase at 50°C?
  5. Does TaqI Methyltransferase require MgCl2?
  6. Does TaqI Methyltransferase require BSA?
  7. What is the activity of TaqI Methyltransferase in other buffers?
  8. What is the molecular weight of TaqI Methyltransferase?
  9. Is TaqI Methyltransferase used in any special techniques?

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Interactive Tools

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • Exonuclease Activity (Radioactivity Release):
    The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
  • Non-Specific DNase Activity (16 hour):
    The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.