| Product Information |
| Omni Klentaq® |
| Part Number |
P7500L |
| Price |
$380 |
| Concentration |
42,000 U/mL |
| Unit Size |
42,000 U |
Product Description:
Omni Klentaq® is an engineered DNA polymerase which lacks both 5'→3' and 3'→5' exonuclease activities and is resistant to the inhibitory effects of complex samples such as blood and soil. This enzyme retains robust PCR function in the presence of greater than 25% whole blood, and a series of mutations provides an effective hot-start function. Omni Klentaq® is licensed by Enzymatics from DNA Polymerase Technologies.
Source of Protein
An E. coli strain carrying a modified recombinant Taq DNA polymerase gene from the strain Thermus aquaticus YT-1.
Supplied in
10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
0.3% Brij-58
50% glycerol
pH 7.4 @ 25°C
Supplied With:
B7500 5X Omni Klentaq®Buffer
5X Omni Klentaq®Buffer (B7500):
250 mM Tris-HCl
80 mM (NH4)2SO4
17.5 mM MgCl2
0.125% Brij 58
pH 9.2 @ 25°C
Unit Definition
1 unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material
in 10 minutes at 75°C.
Contamination Tests:
Single-Stranded Exonuclease Activity
A 50 µL reaction containing 10,000 cpm of a radiolabeled single-stranded DNA substrate and 10 µl of enzyme solution incubated for 4 hours at 37°C resulted in less than 1.0% release of TCA-soluble counts.
Double-Stranded Exonuclease Activity
A 50 µL reaction containing 5,000 cpm of a radiolabeled double-stranded DNA substrate and 10 µl of enzyme solution incubated for 4 hours at 37°C resulted in less than 1.0% release of TCA-soluble counts.
Double-Stranded Endonuclease Activity
A 50 µL reaction containing 0.5 µg of pBR322 DNA and 5 µl of enzyme solution incubated for 4 hours at 37°C resulted in no visually discernible conversion to nicked circular DNA as determined by agarose gel electrophoresis.
E.coli 16S rDNA Contamination Test
Replicate 5 µl samples of enzyme solution were 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. The acceptance criterion for the test is the threshold cycle count (Ct) produced by the average of 3 replicate no template control samples. Based on the correlation between the no template control Ct values, and standard curve data, the detection limit of this assay is <10 copies genome/sample.
References:
1. Kermekchiev, M.B., et al. (2003) Acids Res. 31, 6139-6147.
2. Kermekchiev, M.B. et al. (2009) Nucl. Acids Res., 37 (5):e40 E pub.
3. Zhang Z., et al. (2010) J Mol Diagn. 2, 152-61.
|
| Product Specification* |
| Storage Temperature |
-15 to -25°C |
| TEST: |
SPECIFICATION: |
| Purity (SDS-PAGE) |
>95% |
| Specific Activity |
77,000 – 114,823 U/mg |
| SS Exonuclease |
>350 U <1.0% released |
| DS Exonuclease |
>350 U <1.0% released |
| DS Endonuclease |
>350 U = no conversion |
| E.coli DNA Contamination |
>175 U <10 copies |
* For a detailed summary of assay conditions and data,
refer to the Quality Controls Analysis section below
Quality Control Analysis:
Unit Characterization Assay
Specific activity was 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 ([Omni Klentaq®]f = 0.08-0.0003µg/µL) and added to 50 µL reactions containing 4.0 µg Calf Thymus DNA, 25 mM TAPS (pH 9.3), 50 mM KCl, 1 mM DTT, 4mCi/mL 3H-dTTP and 200 µM 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). Specific activity is determined by dividing the number of measured units by the mass of protein tested. Protein concentration is measured using the Bradford method. The acceptance criterion for specific activity is a range of 77,000-114,823 Units per milligram of protein.
SDS-Page (Physical Purity Assessment) and Specifications
2.0 µL of enzyme solution was loaded on a denaturing 4-20% Tris-Glycine SDS-PAGE gel flanked by a broad-range MW marker and 2.0 µL of a 1:100 dilution of the sample. Following electrophoresis, the gel was stained and the samples compared to determine physical purity. The acceptance criteria for this test requires that the aggregate mass of contaminant bands in the concentrated sample do not exceed 5X the mass of the protein of interest band in the dilute sample, confirming greater than 95% purity of the concentrated sample.
Legal Disclaimers:
Patents
Certain applications in which this product can be used may be covered by patents issued and applicable in the United States and abroad. Purchase of this product does not include a license to perform any patented application; therefore it is the sole responsibility of users of this product to determine whether they may be required to engage a license agreement depending upon the particular application in which the product is used.
This product is sold under license from DNA Polymerase Technology, Inc., U.S. Patent Nos. 7,462,475 and 5,436,149. Omni Klentaq® is a registered trademark of DNA Polymerase Technology, Inc.
|
