Real-time-PCR Master Mix E1
qPCR Master mix with EvaGreen and dUTP
- The Master mix contains dUTP instead of dTTP
- It contains EvaGreen as fluorescent dye
- The qPCR / RT-PCR Mastermix E1 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
-The Master Mix can be used with ROX as reference dye (1x
concentrated)
Applications:
- Detection and quantification of DNA and cDNA targets
- Profiling gene expression
- Microbial detection
- Viral load determination
Compatibility:
BioRad: iCycler, MyiQ, MiQ 2, iQ 5, CFX-96, CFX-384, MJ Opticon, Option2, Chromo4, MiniOpticon
Qiagen: Roto-Gene Q, Roto-Gene3000, Roto-Gene 6000
Eppendorf: Mastercycler realplex
Illumina: Eco RealTime PCR System
Cepheid: SmartCyler
Roche: LIghtCycler 480, LightCycler 2.0
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 dUTP instead of 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, dUTP, EvaGreen, 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
Stability tests / Quality control / Comparison
Manufactured and quality-controlled in accordance with ISO 9001:2000
Usage:
| Components | Volume per reaction | final conc. |
|
2X qPCR / RTD- PCR Master mix E1 |
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
qPCR Master mix with EvaGreen and dUTP
* availibility of sample size may be limited
- verfügbar / avaílable
Deutsche Beschreibung
qPCR Mastermix E1
Datasheet qPCR-Mastermix E1
Material Safety Datasheet
References / Protocols / Notes / Recomendations / Tests
Contact
GeneOn GmbH
Paracelsusstrasse 10
67071 Ludwigshafen am Rhein
Deutschland / Germany
Phone: +49-621-5720 864
Fax: +49-621-5724 462
info@geneon.net
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See Categories
- DNA/RNA Extraction
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GENEON GmbH - Germany - Paracelsusstrasse 10 - 67071 Ludwigshafen - www.geneon.net - info@geneon.net - Tel: 0621-5720864