NIPPON GENE CO., LTD.
URL https://www.nippongene.com/Taq MutS
Code No.

316-04011

Size

50 μg

Concentration

1 μg/μl

Molecular Weight89.3 kDa

SourceThermus aquaticus

Storage Temperature －20℃

Storage Condition
100 mM NaCl, 20 mM Tris-HCl (pH 7.5), 0.1 mM EDTA,
1 mM DTT, 50% Glycerol

Enzyme Reaction Condition
100 mM KCl, 50 mM Tris-HCl (pH 8.5 at 25℃), 20 mM

Binding test:
In 20 μl of the reaction solution, 0.5 μg of theproduct can bind to not less than 50% of 100 ngof a 36 bp synthetic DNA having 1 base deletionat 65°C for 30 min.

Purity:Three μg of the product and 0.5 μg of plasmid
pBR322 were reacted at 37°C for 16 hr and thenagarose gel (0.8% Agarose S) electrophoresis
was performed. The result indicated noincrease of oc-DNA.
Three μg of the product and 0.5 μg of λDNA
were reacted at 37°C for 16 hr and then agarosegel (0.8% Agarose S) electrophoresis wasperformed. The result indicated no degradationof λDNA.
Three μg of the product and 2 μg of substrate
RNA were reacted at 37°C for 16 hr and thenagarose gel (2% Agarose S) electrophoresis wasperformed. The result indicated no degradationof RNA.

MgCl2, 0.1 mM EDTA, 1 mM DTT, 2% Glycerol
Reaction Temperature： 65℃
References■ The attached exclusive buffer for the enzyme
reaction（Store at -20℃）

1) Biswas, I. and Hsieh, P.: J. Biol. Chem., 271 (9),・10×Taq MutS Buffer2) Biswas, I. and Hsieh, P.: J. Biol. Chem., 272 (20),1 ml

It is 10-fold concentrated compared to theenzyme reaction condition.

5040-5048 (1996)
13355-13364 (1997)
3) Takamatsu, S., Kato, R. and Kuramitsu, S.:
Nucleic Acids Res., 24 (4), 640-647 (1996)
4) Whitehouse, A., Deeble, J., Parmar, R., Taylor, G. R.,
Markham, A. F. and Meredith, D. M.: Biochem.
Biophys. Res. Commun., 233, 834-837 (1997)
5) Lishanski, A., Ostrander, E. A. and Rine, J.:
Proc. Natl. Acad. Sci. USA, 91, 2674-2678 (1994)
6) Wagner, R., Debbie, P. and Radman, M.:
Nucleic Acids Res., 23 (19), 3944-3948 (1995)

For research use only120510KH

NIPPON GENE CO., LTD.
URL https://www.nippongene.com/Electrophoresis Running buffer

Example of use

・ 1×TAE

Test for detecting the deleted region using PCR

・ 0.1 mM MgCl2products↓

A plasmid having a mutated sequence (M: mutant) in

Stain the gel with SYBR® Gold

which 2 nucleotides are deleted as compared to thenormal sequence (N: normal) was produced.
Respective PCRs were performed using N or M singly,↓or a mixture of N and M as templates. After completing
60 bp

PCR, the samples were denatured and annealed, andreacted by directly adding Taq MutS (1 μg) at 65 °C forN

M N/M

100 bp200 bp

NN

M N/M

M

30 min, and then subjected to electrophoresis. PCR←Taq MutS+ DNA

was performed so that the lengths of the amplified
products were 3 kinds of 60 bp, 100 bp and 200 bp, and←DNA

the deleted region was detected in each of the products.
Template DNA (N/M, N or M)
1 μl
10×Gene Taq Universal Buffer
5 μl
dNTP Mixture (2.5 mmol/l each)
4 μl
Primer-forward (20 pmol/μl)
1 μl
Primer-reverse (20 pmol/μl)
1 μl
Gene Taq (5 units/μl)
0.5 μlH2O37.5 μlTotal
50 μl
↓
94℃
1 min.
94℃
10 sec.
60℃
15 sec.
25 cycles
72℃
20 sec.
94℃
5 min. (denaturation of PCR products)
72℃
30 min. (re-annealing)
4℃

N/M( For more details on experimental examples and thelike, see the website of Nippon Gene.)↓
PCR products 10 μl ＋ Taq MutS 1 μl (1 μg/μl)↓65℃, 30 min.
↓＋ 2 μl Loading buffer(0.02% Bromophenol blue, 0.02% Xylene cyanol FF,
50% Glycerol, 200 mM Tris-HCl pH 7.5)
↓NIPPON GENE CO., LTD.
2-7-18, TOIYA-MACHI, TOYAMA 930-0834 JAPANTel+81-76-451-6548
URL
https://www.nippongene.com/Electrophoresis in polyacrylamide gradient gel
Polyacrylamide gradient gel
・ 4－10％ native polyacrylamide
・ 1×TAE

This product is sold as a reagent (for research) and is
not for pharmaceutical use.

・ 0.1 mM MgCl2

For research use only
120510KH