Features:
- Enzyme with hot start capability increases reaction specificity and
sensitivity, optimized
- DFS-Taq PLUS DNA polymerase activation requires not more than 5
min heating
- High selectivity and reaction yield
- The mix is colored for easy pipetting
- Reduced preparation time
Applications:
- Real-time PCR with oligonucleotide probes
- Conventional PCR
- High-throughput PCR
- Genotyping
- For detection of bacterial DNA
Platforms:
(ABI) 7000, 7300, 7700, 7900, 7900HT and StepOnePlus
Description
Bio-Star PCR-Mastermix (2×) HIGH-ROX for probes is developed for quantitative real-time PCR with fluorescent probes. The Mastermix contains all components, except template and primers, for successful PCR.
- The mix is optimized for efficient and reproducible hot-start real-time
PCR of genomic, plasmid and viral DNA samples.
- The solution contains substances that increase half-life and
processivity of DFS-Taq PLUS DNA polymerase blocked by MAB amd
by enhancing its stability during PCR.
- It includes components that influence primer annealing temperature
and characteristics of template melting thus enabling to increase the
specificity of PCR and use templates with complicated structure.
- DFS-Taq Plus DNA polymerase is inactive at room temperature
because of monoclonal antibodies.
Components and Mixture
2X Bio-Star PCR-Master mix HIGH-ROX for probes contains:
- 100 mM Tris-НCl (рН 8.5 at 25 °С), 100 mM KCl, 0.4 mM each of
dNTP, 10 mM MgCl2, 0.1 U/µl DFS-Taq DNA Polymerase, 0.025%
Tween 20, 900 nM ROX fluorescent dye stabilizers and enhancers
- Tube of MgCl2 100 mM
- Water Mol.Bio Grade 2 x 1,25 ml
Storage and
transportation: at -20 °С; not more
than 50 thawing-freezing cycles. Shipping with blue ice or at room temperature
Storage
terms: up to 24 mounts
Exodeoxyribonuclease activity: DNA was stable after incubation of 1 µg fragment of phage lambda DNA in the presence of 25 µl of Bio-Star qPCR Mastermix (2×) in 50
µl reaction solution at 37 °C and 70 °C for 4 h.
Test for bacterial DNA contamination
Test for Hot -Start ( we test activity with or without MAB at various temperatures
Exodeoxyribonuclease activity
DNA was stable after incubation of 1 μg fragment of phage lambda DNA in the presence of 25 μl of BioMaster HS-qPCR (2×) in 50 μl reaction solution at 37 °C and 70 °C for 4 h.
Ribonuclease activity
Absence of ribonuclease activity was confirmed after incubation of 1 μg of 5’-[P32]-labeled RNA fragment in the presence of 25 μl of BioMaster HS-qPCR (2×) in 50 μl reaction solution at 37 °C for 4 h.
Stability:
One month at room temperature does not reduce PCR efficiency
1. Defrost the reaction mixture and stir thoroughly.
2. Add the following components into the thin-wall PCR tubes considering the final volume of a reaction mixture equal to 50 μl:
Component | Volume | Final concentration |
2x Mastermix | 25 µl | 1x |
Forward Primer | variable | 0.1 - 600 nM |
Reverse Primer | variable | 0.1 - 600 nM |
DNA Template | variable | 10 pg - 1 µg |
Sterile Water | up to 50 µl |
3. Gently vortex and remove droplets by centrifugation.
4. Perform PCR using temperature conditions recommended below:
Step |
Temp. °C |
Incubation time
|
Number of Cycles |
Preliminary denaturation |
95 | 3-7 min | 1 |
Denaturation |
95 | 15 sec | 25-40 |
Annealing |
50-68 |
10-30 sec |
25-40 |
Elongation | 58-72 | 30-60 sec | 25-40 |
or alternatively:
Step |
Temp. °C |
Incubation time |
Number of Cycles |
Preliminary denaturation |
95 | 3-7 min | |
Denaturation |
95 | 15 sec | 30-50 |
Anealing/Elongation |
50-68 | 1 min | 30-50 |
5. The PCR results are displayed as amplification curve
Applications:
- Real-time PCR with oligonucleotide probes
- Conventional PCR
- High-throughput PCR
- Genotyping
- For detection of bacterial DNA
Final price excl. shipping costs3
PCR MASTERMIX SELECTION TABLE
PCR Mastermixes with dTTP and SybrGreen
BioStar 2 PCR mastermix with SYBRGreen, blue dye, no
rox
BioStar 3 PCR mastermix with SYBRGreen, 60 nm LOW-ROX
BioStar 4 PCR mastermix with SYBRGreen, 900 nm HIGH-ROX
PCR Mastermixes with dTTP for probes
BioStar 5 PCR mastermix for probes
BioStar 6 PCR mastermix for probes 60 mM LOW-ROX
BioStar 7 PCR mastermix for probes 900 mM HIGH-ROX
PCR Master Mixes with Evagreen and dUTP
PCR Master Mix for qPCR EVA 1 dUTP+ EvaGreen
PCR Master Mix for qPCR EVA 2 +UNG
PCR Master Mix for qPCR EVA3 +ROX
PCR Master Mix for qPCR EVA4 +ROX+UNG
PCR Master Mix for qPCR EVA5 +low ROX
PCR Master Mix for qPCR E6 +low ROX+UNG
PCR Master Mixes for probes and dUTP
qPCR Master Mix DLP1 Exclusiv +dUTP
qPCR Master Mix DLP2 Exclusiv +dUTP+UDG
qPCR Master Mix DLP3/DLP5 Exclusiv ROX (500 nM, 100nM)
qPCR Master Mix DLP4/DLP6 Exclusiv +ROX 500nM or 100 nM+UDG
Bio-Star Mastermix HIGH-ROX für "fluorescent probes"