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商品編號


P7620L



品牌


Enzymatics



公司


Enzymatics,inc



公司分類


New Products



Size

5,000 U

商品信息

TaqIT


Product Description

TaqIT is an exonuclease deficient derivative of Taq DNA polymerase. TaqIT lacks the first 280 amino acids of native Taq polymerase that contain the 5′-3′ exonuclease domain. This deletion makes TaqIT slightly more
Thermo
stable and has slightly greater fidelity than full length Taq. Like Taq polymerase, TaqIT has no inherent 3′-5′ exonuclease activity.

Source of Protein
A recombinant
E. coli
strain carrying the TaqIT gene from the
Thermo
philic organism Thermus Aquaticus YT-1.

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.

Molecular weight
62,407 Daltons



Quality Control Analysis

Unit Activity
is measured using a 2-fold serial dilution method. Dilutions of enzyme were made in a reduced-glycerol (5%) containing Taq-B DNA Polymerase storage solution and added to 50 ?L reactions containing Calf Thymus DNA, 25 mM TAPS (pH 9.3), 50 mM KCl, 1 mM DTT, 3H-dTTP and 100 ?M dNTPs. Reactions were incubated 10 minutes at 75°C, plunged on ice, and analyzed using the method of Sambrook and Russell (Molecular Cloning, v3, 2001, pp. A8.25-A8.26).

Protein Concentration
is determined by OD
280
absorbance.

Physical Purity
is evaluated by SDS-PAGE of concentrated and diluted enzyme solutions followed by silver stain detection. Purity is assessed by comparing the aggregate mass of contaminant bands in the concentrated sample to the mass of the protein of interest band in the diluted sample.

Single-Stranded Exonuclease
is determined in a 50 ?L reaction containing a r
ADI
olabeled single-stranded DNA substrate and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

Double-Stranded Exonuclease
is determined in a 50 ?l reaction containing a r
ADI
olabeled double-stranded DNA substrate and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

Double-Stranded Endonuclease
is determined in a 50 ?L reaction containing 0.5 ?g of plasmid DNA and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

E.coli
16S rDNA Contamination
is evaluated using 5 ?L
r
eplicate samples of enzyme solution denatured and screened in a TaqMan qPCR assay for the presence of contaminating
E.coli
genomic DNA using oligonucleotide primers corresponding to the 16S rRNA locus.

Supplied in:
20mM Tris-HCl
222mM (NH
4
)
2
SO
4
10mM 2-mercaptoethanol
0.1 mM EDTA
0.1% Brij 58
50% glycerol
pH 8.6 @ 25°C


Supplied with:

10X TaqIT Reaction Buffer (B7620):
500mM Tris-HCl
160 mM (NH
4
)
2
SO
4
35mM MgCl
2
0.25% Brij 58
pH 9.2 @ 25?C


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Product Information



TaqIT


Part Number
P7620L
Price
$503
Concentration
50,000 U/mL
Unit Size
5,000 U


SDS

Available on request








Product Specification*


Storage Temperature
-25?C to -15?C


Test

Units Tested

Specification


SDS Purity
n/a
>99%
Specific Activity
n/a
42,000 U/mg
SS Exonuclease
500
<1.0% Released
DS Exonuclease
500
<1.0% Released
DS Endonuclease
500
No Conversion
E.coli
DNA Contamination
500
<10 copies





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

References


The fidelity of Taq polymerase catalyzing PCR is improved by an N-terminal deletion. Barnes WM. Gene. 1992. 112(1):29-35.

Comparative thermal denaturation of Thermus aquaticus and Escherichia coli type 1 DNA polymerases. Karantzeni I, Ruiz C, Liu CC, Licata VJ. Biochem J. 2003 374(Pt 3):785-92.




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.