Features:
- The Master mix contains dUTP instead of dTTP
- The Mix contains ROX (500nM; LOW-ROX-version 100mM) as passive
Reference dye (it provides a baseline in multiplex reactions)
- It contains EvaGreen as fluorescent dye
- The qPCR / RTD-PCR Master mix E3 is ready-to-use and is optimized
for high specificity and sensitivity because of optimized reaction buffer
- easy to use because ready-to-use Master Mix
Compatibility:
500 nM Version: ABI: 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne, StepOne plus
100 nM Version: 7500 System, Stratagene Mx35005P, Mx4000 and Mx3000P, ViiA 7, QuantStudio
Applications:
- Detection and quantification of DNA and cDNA targets
- Profiling gene expression
- Microbial detection
- Viral load determination
Description:
The PCR-Master Mix contains all reagents required for qPCR (except template and primer) in a premixed 2x concentrated ready-to-use solution. The high
specificity and sensitivity of the mix is achieved by an optimized hot-start polymerase. Its activity is blocked at ambient temperature and switched on automatically at the onset of the initial
denaturation. The thermal activation prevents the extension of non-specifically annealed primers and primer-dimer formations at low temperatures during PCR setup.
The PCR-mix for qPCR offers dUTP instead of dTTP to prevent carry-over contaminations of DNA from previous PCR reactions.
Concentration: The Mastermix is 2x concentrated
List of components qPCR / RTD-PCR Master mix:
Hot-Start Polymerase for qPCR, dATP, dCTP, dGTP, dUTP, EvaGreen, ROX, optimized reaction buffer with KCl and MgCl2,
stabilizers and enhancers, PCR-grade water
Transportation: with blue ice
Storage: at 4°C for 3 months, at -20°C for more than 12 months,
Note: protect from Light
Manufactured and quality-controlled in accordance with ISO 9001:2000
Components | Volume per reaction | final conc. |
2X qPCR / RTD- PCR Master mix E3 |
25 µl | 1x |
Up-stream primer (10 µM stock) |
1,5 µl (range: 0,5-2.5 µl) | 300 nM |
Down-stream primer (10µM stock) |
1,5 µl (range: 0.5-2,5 µl | 300 nM |
Template DNA |
5 µl (0.1-15 ng/ml plasmid DNA) (1-10 µg/ml genomic DNA) |
< 500ng DNA |
Sterile dest. Water (included) |
up to 50 µl total reaction volume |
- vortex all solutions carefully before using and before PCR
- may you add the enzyme mix after Template DNA
- an individual optimization of annealing temperature may be
necessary for new combinations of primers and Template DNA
General Thermo-Cycler protocol for qPCR / RTD-PCR Master mix:
Note: working with EvaGreen just select the optical setting for FAM or SYBR Green at the cycler
Step | Time | Temperature |
Initial denaturation | 1-3 min | 95°C |
30-40 Cycles: Denaturation Annealing Extension |
15-30 sec 30-65 sec 30 sec (per 500bp) |
95°C 55-65°C 72-75°C |
Note:
an individual optimization of annealing temperature may be necessary for new combinations of primers and Template DNA
The PCR-Master Mix contains all reagents required for qPCR (except template and primer) in a premixed 2x concentrated ready-to-use solution. The high specificity and sensitivity of the mix is achieved by an optimized hot-start polymerase. Its activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation.
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