商品编号


P7590L



品牌


Enzymatics



公司


Enzymatics,inc



公司分类


Ultrapure Polymerases



Size

500 U

商品信息

Phoenix? Hot Start Taq DNA Polymerase


Product Description

Phoenix? Hot Start Taq DNA Polymerase is a recombinant,
Thermo
st
ABI
le Taq DNA polymerase complexed with a
Thermo
l
ABI
le, neutralizing antibody that blocks the 5′→3′ polymerase activity prior to the initial DNA denaturation step of PCR (1,2). Such antibody-mediated Hot-Start cap
ABI
lity enhances the overall specificity, sensitivity and yield of the PCR by reducing nonspecific amplification and primer-dimer formation prior to PCR cycling, and allows the convenience of reaction set up at room temperature. When the temperature of the PCR reaction mix reaches ≥94°C during the initial DNA denaturing step of PCR cycling, activity of the Taq DNA polymerase is fully restored. Phoenix Hot Start Taq DNA Polymerase, like standard Taq DNA polymerase, also has 5′→3′ exonuclease activity, but lacks any detectable 3′→5′ exonuclease activity.

Source of Protein
A recombinant?
E. coli
?strain carrying the Taq DNA polymerase gene from the
Thermo
philic organism Thermus Aquaticus YT-1 complexed with a monoclonal antibody derived from murine cell culture.

Supplied in
20 mM Tris-HCl
100 mM NaCl
0.1 mM EDTA
St
ABI
lizer
50% Glycerol
pH 7.5 @ 25°C

Supplied With:
5X Phoenix Hot Start Taq Reaction Buffer (B7590)
5X Phoenix Hot Start Taq GC Reaction Buffer (B7591)

Unit Definition
1 unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C.



Quality Control Analysis

Functional Assay
Phoenix Hot Start Taq DNA polymerase was tested for its
ABI
lity to amplify DNA targets by PCR in reaction buffers B7590 and B7591 following a 24 hour incubation at room temperature. After heat activation and PCR amplification the samples resulted in vis
IBL
e single band amplicons as determined by agarose gel electrophoresis.

Taq Inhibition
This assay measured the residual activity of Phoenix Hot Start Taq DNA polymerase in the absence of a ≥ 94°C heat activation step. Phoenix Hot Start Taq DNA polymerase and Taq DNA Polymerase (Taq DNA Polymerase without Taq Antibody) were incubated in the presence of Calf Thymus DNA, 50 ?M?
3
H-dTTP, 100 ?M dNTPs and 1X reaction buffer B7590 or B7591. After a 24 hour incubation at room temperature the samples were analyzed for total 3H-dTTP counts incorporated using the method of Sambrook and Russell (Molecular Cloning, v3, 2001, pp. A8.25-A8.26).



Contamination Tests

Single-Stranded Exonuclease Activity
R
ADI
olabeled single-stranded DNA substrate incubated with enzyme solution for 4 hours at 37°C resulted in less than 5% release of TCA-soluble counts.

Double-Stranded Exonuclease Activity
R
ADI
olabeled double-stranded DNA substrate incubated with enzyme solution for 4 hours at 37°C resulted in less than 1% release of TCA-soluble counts.

Double-Stranded Endonuclease Activity
A reaction containing plasmid DNA incubated with enzyme solution for 4 hours at 37°C resulted in no visually discern
IBL
e conversion to nicked circular DNA as determined by agarose gel electrophoresis.


View PDF Poster Instructions FAQ


Product Information



Phoenix? Hot Start Taq DNA Polymerase


Part Number
P7590L
Price
$213
Concentration
5,000 U/ml
Unit Size
500 U


SDS

Available on request








Product Specification*


Storage Temperature
-25 to -15°C


Test

Specification


Specific Activity, Taq DNA Polymerase (P7580)
74,625 U/mg
SS Exonuclease
50 U < 10%
DS Exonuclease
50 U < 1%
DS Endonuclease
50 U = No conversion
Taq Inhibition
Pass
Functional Assay
Functional with buffers B7590 and B7591





* For a detailed summary of assay conditions and data, refer to the Quality Controls Analysis section.

Additional Downloads

Product Note

References


Chou, Q. et al. (1992) Nucleic Acids Research 20(7), 1717-1723.

Sharkey, D. et al. (1994), Nature Biotechnology, 12, 506 – 509.




Limitations of Use
This product was developed, manufactured, and sold for in vitro use only. The product is not suitable for admi
NIST
ration to humans or animals. SDS sheets relevant to this product are available upon request.