商品编号


A1905B



品牌


Lucigen



公司


Lucigen



公司分类


Modifying Enzymes



Size

5 ml

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.