Description:
The 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 mix contains dTTP and allows an UNG (Uracil-N-Glycosylase) treatment at the onset of thermal cycling to prevent carry-over contaminations of DNA from previous PCR reactions.
Concentration: The Master mix is 2x concentrated
List of components:
Hot-Start Polymerase for qPCR, dATP, dCTP, dGTP, dTTP, EvaGreen, optimized reaction buffer with KCl and (NH4)2SO4, 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
Usage:
Components | Volume per reaction | final conc. |
2X qPCR- PCR Master mix E1 |
25 µl | 1x |
Up-stream
primer (10 |
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. |
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
Features:
- The PCR Master mix contains dTTP
- It contains EvaGreen as fluorescent dye
- The qPCR / RT-PCR Mastermix EVA2 is ready-to-use and is optimized for high specificity and sensitivity
because of optimized reaction buffer
- easy to us because ready-to-use Master Mix
- The Master Mix can be used with ROX as reference dye (1x concentrated)
Final price3
qPCR Mastermix E1