Description
The ExcelTaq™ Blood Direct DNA Polymerase is designed for amplifying targeted DNA directly from whole blood, eliminating the need for a lengthy DNA isolation process. ExcelTaq™ Blood Direct DNA Polymerase is highly tolerant in the presence of PCR interfering/ inhibiting substances in blood, such as IgG, hemoglobin, and lactoferrin. ExcelTaq™ Blood Direct DNA Polymerase is compatible with most anticoagulants, such as citrate, EDTA, and heparin (Fig. 1). The ExcelTaq™ Blood Direct DNA Polymerase includes a pair of Positive Control Primers (CCR5) that are compatible with primate blood samples.
Features
5'→3' DNA polymerase activity
No detectable 3'→5' exonuclease (proofreading) activity
Generates PCR products with 3'-dA overhangs
Compatible with most anticoagulants
Applications
Direct amplification of DNA from blood samples
High throughput screening without DNA purification
Suitable for multiplex PCR
Storage
-20°C for 24 months
4℃ for 6 months
Compatible with most anticoagulants
ExcelTaq™ Blood Direct DNA Polymerase amplified 200 bp from differently treated blood using CCR5 specific primers. (M: DM2100)
Contents
|
Storage Buffer
20 mM Tris-HCl pH 8.0, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizer, 50% (v/v) glycerol
CCR5 control primer sequence
CCR5-F: 5'-CTCCCAGGAATCATCTTTACC-3'
CCR5-R: 5'-TCATTTCGACACCGAAGCAG-3'
Storage
-20°C for 24 months
4℃ for 6 months
Recommended PCR Condition
Blood |
3 μl |
Forward primer |
0.1 – 0.5 µM |
Reverse primer |
0.1 – 0.5 µM |
5X Blood Direct Buffer |
10 μl |
dNTPs |
0.2 mM (each) |
Blood Direct DNA Polymerase |
0.25 μl (1.25 U) |
ddH2O |
to 50 μl |
Total volume |
50 µl |
Note: Mix well before PCR Reaction
Recommended PCR Program
Steps |
Temp. |
Time |
Cycles |
Template denature |
94°C |
3 min |
1 |
Denature |
94°C |
30 sec |
30-40 |
Annealing |
50-68°C* |
30 sec |
|
Extension |
72°C |
60 sec/kb |
|
Final extension |
72°C |
3 min |
1 |
*Optimal PCR condition varies according to primers’ thermodynamic properties.
Post amplification analysis
Centrifuge PCR reaction mixture at 1000 × g for 1 ~ 3 minutes. Load only the clear supernatant of the PCR products for gel electrophoresis analysis.
[TP2100] ExcelTaq™ 5X Blood Direct PCR Master Mix Kit
5'→3' DNA polymerase activity
None detectable 3'→5' exonuclease (proofreading) activity
Generates PCR products with 3'-dA overhangs
High throughput PCR
Execute PCR directly from blood samples
High reproducibility, less pipetting errors
Compatible with most anticoagulants
[TP5000] ExcelTaq™ Hot Start II DNA Polymerase
Aptamer-based hot start PCR
Reversible enzyme inactivation
Omits extra enzyme activation step
Convenient for room temperature PCR set-up
High yield and specificity of target amplicons
Wide range of amplicon length (up to 10 kb)
High sensitivity (as low as 1 fg of plasmid)
Aptamer-based hot start PCR
Reversible enzyme inactivation
Omits extra enzyme activation step
Convenient for room temperature PCR set-up
High yield and specificity of target amplicons
Wide range of amplicon length (up to 10 kb)
High sensitivity (as low as 1 fg of plasmid)
[TQ1200] ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX)
High Stability
Fast Hot Start
High Sensitivity
Low Background / High Specificity
Suitable for Fast Program
Smart Blue Contrast Dye
[TP1200] ExcelTaq™ 5X PCR Master Dye Mix
5’→3’ DNA polymerase activity
No detectable 3'→5' exonuclease (proofreading) activity
Generates PCR products with 3'-dA overhangs
High yield PCR
High reproducibility
Reduced pipetting errors
Includes tracking dye for direct loading after PCR