Taq MutS

Product	Size	Code No.
Taq MutS	50µg	316-04011

 

Taq MutS is an enzyme that acts in the DNA repair system, recognizes mismatch base pairs of DNA and binds thereto.
Being derived from Thermus aquaticus, Taq MutS is heat-stable (0-75°C) and stays active and binds to DNA even at high temperatures.
Since Taq MutS efficiently binds to 1-4 bases deletion (or insertion) and mismatch base pairs of GT, CT and AG, it is very useful for detecting these mutations. Using this specific binding activity, mutations can be detected in polyacrylamide gels or on a solid phase.
In this product, 1 ml of 10 x Taq MutS Buffer is attached as the exclusive buffer for the enzyme reaction.

Source	Thermus aquaticus
Molecular Weight (M.W.)	89.3 kDa
Concentration	1 µg/µl
Storage Condition	20mM Tris-HCl(pH 7.5), 100mM NaCl, 0.1mM EDTA, 1mM DTT, 50%(v/v) Glycerol
Enzyme Reaction Condition	100mM KCl, 50mM Tris-HCl(pH 8.5), 20mM MgCl2, 0.1mM EDTA, 1mM DTT, 2% Glycerol, 65℃

The attached exclusive buffer for the enzyme reaction is 10-fold concentrated compared to the enzyme reaction condition.

−20℃(should not be frozen)

Gel-shift assay using a synthetic DNA of 36 bp.
The difference of the binding activity of Taq MutS to 1-4 base deletion (Lane 2-5) and 8 kinds of mismatches (Lane 6-13) was confirmed.

	Lane1 AT(control) Lane8 AG Lane2 Δ1 Lane9 TT Lane3 Δ2 Lane10 AC Lane4 Δ3 Lane11 AA Lane5 Δ4 Lane12 CC Lane6 GT Lane13 GG Lane7 CT Lane14 AT(control)
6% non-denaturing polyacrylamide gel SYBR® Gold staining

In 20 µl of the reaction solution, 0.5 µg of the product can bind to not less than 50% of 100 ng of a 36 bp synthetic DNA having 1 base deletion at 65°C for 30 min.

• Three µg of the product and 0.5 µg of plasmid pBR322 were reacted at 37°C for 16 hr and then agarose gel (0.8% Agarose S) electrophoresis was performed. The result indicated no increase of oc-DNA.
• Three µg of the product and 0.5 µg of λDNA were reacted at 37°C for 16 hr and then agarose gel (0.8% Agarose S) electrophoresis was performed. The result indicated no degradation of λDNA.
• Three µg of the product and 2 µg of substrate RNA were reacted at 37°C for 16 hr and then agarose gel (2% Agarose S) electrophoresis was performed. The result indicated no degradation of RNA.

1. Biswas, I. and Hsieh, P. : J. Biol. Chem., 271（9）, 5040（1996）
2. Biswas, I. and Hsieh, P. : J. Biol. Chem., 272（20）, 13355（1997）
3. Takamatsu, S., Kato, R. and Kuramitsu, S. : Nucleic Acids Res., 24（4）, 640（1996）
4. Whitehouse, A., Deeble, J., Parmar, R., Taylor, G. R., Markham, A. F. and Meredith, D. M. : Biochem. Biophys. Res. Commun., 233, 834（1997）
5. Lishanski, A., Ostrander, E. A. and Rine, J. : Proc. Natl. Acad. Sci. USA, 91, 2674（1994）
6. Wagner, R., Debbie, P. and Radman, M. : Nucleic Acids Res., 23（19）, 3944（1995）

Taq MutS manual

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