商品编号


A3210K



品牌


Lucigen



公司


Lucigen



公司分类


Modifying Enzymes



Size

10,000 Units

商品信息

Highly
Thermo
stable DNA ligase for applications where ligation at high temperature is beneficial

High
Thermo
st
ABI
lity allows ligation using high-stringency hybridization conditions

High specificity and stringency permits sensitive detection of SNPs
11





Applications

Ligation amplification (Ligase Chain Reaction, LCR)
1-7

Repeat Expansion Detection (RED)
7

Simultaneous mutagenesis of multiple sites
8,9

Other ligation-based detection methods

Ampligase?
Thermo
stable DNA Ligase catalyzes NAD-dependent ligation of adjacent 3?-hydroxylated and 5?-phosphorylated termini in duplex DNA structures that are stable at high temperatures. Derived from a
Thermo
philic bacterium, the enzyme is stable and active at much higher temperatures than conventional DNA ligases. Its half-life is 48 hours at 65°C and greater than 1?hour at 95°C. Ampligase DNA Ligase has been shown to be active for at least 500 thermal cycles (94°C/80°C) or 16 hours of cycling.
10
This exceptional
Thermo
st
ABI
lity permits extremely high hybridization stringency and ligation specificity. Ampligase DNA Ligase has no detectable activity in ligating blunt-ended DNA and has no activity on RNA or RNA:DNA hybrids.

Unit Definition:
One unit of Ampligase DNA Ligase catalyzes the ligation of 50% of the
cos
sites in 1 ?g of lam
BD
a DNA in 1 minute at 45°C in 1X Ampligase Reaction Buffer.

Note:
One unit of Ampligase DNA Ligase is equivalent to at least 15 of the "cohesive end units" or "nick ligation units" defined elsewhere.
4

Storage Buffer:
50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton? X-100.

Ampligase 10X Reaction Buffer:
200 mM Tris-HCl (pH 8.3), 250 mM KCl, 100 mM MgCl
2
, 5 mM NAD, and 0.1% Triton? X-100.

Quality Control:
Ampligase
Thermo
stable DNA Ligase is assayed for the absence of blunt-end ligation activity by ligating
Sma
I-digested bacteriophage lam
BD
a DNA at 45°C; ligation occurs only at
cos
sites as determined by gel electrophoresis. Ampligase DNA Ligase is free of detectable exo- and endonuclease and RNase activities.

References

Landegren, U.
et al.
(1988)
Science

242
, 229.

Wu, D.Y. and Wallace, R.B. (1989)
Genomics
4
, 560.

Barany, F. (1991)
Proc. Natl. Acad. Sci.
USA
88
, 189.

Birkenmeyer, L. and Armstrong, A.S. (1992)
J. Clin. Micro.
30
, 3089.

Dille, B.J.
et al.
(1993)
J. Clin. Micro.

31
, 729

Nakazawa, H.
et al.
(1994)
Proc. Natl. Acad. Sci.
USA
91
, 360.

Sirugo, G. and Kidd, K. (1995)
Epicentre Forum

2
(3), 1.

Moore, D. and Michael, S. (1995)
Epicentre Forum

2
(4), 4.

Rouwendal, G.J.
et al.
(1993)
BioTechniques

15
, 68.

Schalling, M.
et al.
(1993)
Nature Genetics

4
, 135.

Hardenbol, P.?
et al.
?(2003)?
Nature Biotechnology
?
21
, 673.


ORDER INFORMATION
Ampligase? is provided at a standard concentration of 5 U/μl, with the exception of catalog numbers A0102K and A110K (100 U/ μl).

Ampligase DNA Ligase Kit contains: Ampligase DNA Ligase, 10X Reaction Buffer, and Control DNA. Ampligase Enzyme & Buffer contain: Ampligase DNA Ligase and 10X Reaction Buffer (25 μl Buffer for each 50 U enzyme).

One unit of Ampligase? is equal to as many as 15 units of other
Thermo
stable DNA ligases. Please compare competitive unit definitions.