Description
The ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is designed for amplifying targeted DNA directly from whole blood, eliminating the need for a lengthy DNA isolation process. ExcelTaq™ Blood Direct PCR Master Mix Kit is a unique blend of 5X concentrated mixture containing all the essential components of a PCR reaction. The ExcelTaq™ Blood Direct PCR Master Mix Kit also contains the PCR-friendly loading and tracking dye solution Orange G. The inclusion of Orange G tracking dye allows for a more convenient post-PCR gel electrophoresis analysis. ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is capable of tolerating the presence of PCR interfering/ inhibiting substances in blood. The ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is ideal for high-throughput screening of blood samples for high reproducibility. ExcelTaq™ 5X Blood Direct DNA PCR Master Mix Kit 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
High throughput PCR
High yield PCR
High reproducibility, with reduced pipetting errors
Compatible with most anticoagulants
Applications
Direct amplification of DNA from blood samples
High throughput screening without DNA purification
Suitable for multiplex PCR
Storage
Caution: Avoid Multiple Freeze/Thaw Cycles
4°C for 6 months
-20°C for 24 months
Compatible with most anticoagulants
ExcelTaq™ 5X Blood Direct PCR Master Mix amplified 200 bp from differently treated blood using CCR5 specific primers. (M: DM2100)
Contents
Component |
Volume |
5X Blood Direct PCR Master Mix |
1 ml x 2 |
Positive Control Primers (10 μM, each) |
25 μl |
CCR5 control primer sequence
CCR5-F: 5'-CTCCCAGGAATCATCTTTACC-3'
CCR5-R: 5'-TCATTTCGACACCGAAGCAG-3'
Storage
Caution: Avoid Multiple Freeze/Thaw Cycles
4°C for 6 months
-20°C for 24 months
Recommended PCR Condition
Blood |
3 μl |
Forward primer |
0.1 – 0.5 µM |
Reverse primer |
0.1 – 0.5 µM |
5X Blood Direct PCR Master Mix |
10 μl |
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.
[TP2000] ExcelTaq™ Blood Direct DNA Polymerase
High yield
Thermostable, up to 50°C, during first strand synthesis
High processivity, generating cDNA up to 8 kb
Reduced RNase H ribonuclease activity
[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)
[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