DFS-Gen-Taq Mastermix (2X)
| Catalog # | Pack size | Price, Euro | SHOP |
| 101605 | 100 rcs (2x1,25ml) | 49,00 | Add to cart |
| 101625 | 5 x 100 rcs (10x1,25ml) | 220,00 | Add to cart |
Datasheet: DFS-Gen-Taq Master Mix (A)
Description:
2x PCR Master Mix is an optimized ready-to-use PCR mixture of Taq DNA Polymerase, PCR buffer, MgCl2 and dNTPs. 2x PCR Master Mix contains all components for PCR, except DNA template and primers. The mixture contains DFS-Taq DNA Polymerase - Taq DNA Polymerase without traces of E.coli DNA, but with the increased ability to detect low-copy number genes (Bioron GmbH, cat# 101005, 101025, 101100, 101500). Due to the usage of DFS-Taq DNA polymerase the mixture can be recommended not only for all regular applications, but also for templates which contains E.coli DNA sequences.
2x PCR Master Mix Composition
- Taq DNA Polymerase (recombinant) in reaction buffer: 0.1 units/ µl
- 32 mM (NH4)2SO4
- 130 mM Tris-HCl, pH 8.8 at 25oC
- 0,02% Tween-20
- 5,5 mM MgCl2
- dNTPs (dATP, dCTP, dGTP, dTTP): 0.4 mM of each
Performance and purity tests:
Tested for the absence of endodeoxyribonucleases and exodeoxyribonucleases. Performance is tested on single-copy genes of human and mouse DNA.
Endodeoxyribonuclease Assay:
No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 25 µl of 2x PCR Master Mix with 1 µg of pUC19 DNA in 50 µl for 4 hours neither at 37°C nor at 70oC.
Storage:
MasterMix should be stored at -20oC. Repeated (up to 10 times) freezing - defreezing do not cause the reduction in PCR performance, storage at +4oC for 1 week do not cause the reduction in PCR performance.
Protocol: It’s strongly recommended to settle-up all reactions in ice to avoid formation of unspecific products or primer-dimers formation.
Add in a thin walled PCR tube:
| 50 µl reaction volume | 50 µl reaction volume |
| component | volume | final conc. | volume | final conc. |
| 2x PCR Master mix | 25 µl | 1x | 12.5 µl | 1x |
| MgCl2 | variable | variable | variable | variable |
| forward primer | variable | 0.1 - 1 µl | variable | 0.1 - 1 µl |
| reverase primer | variable | 0.1 - 1 µl | variable | 0.1 - 1 µl |
| Template DNA | variable | 10 pg-1µg | variable | 10 pg-1µg |
| Sterile Deionized water | up to 50 µl | - | up to 25 µl | - |
• Gently vortex the sample and briefly centrifuge to collect all drops to the bottom of the tube.
• Overlay the sample with mineral oil or add an appropriate amount of wax if the thermal cycler is not equipped with a heated lid.
• Place the samples in a thermocycler and start the optimal PCR program.
If MgCl2 is not added to the reaction mixture, final concentration of MgCl2 in the reaction mixture will be 2,75mM. Addition of 1 µl of 100 mM MgCl2 into the mixture results in concentration of MgCl2 4,75mM in the reaction mixture for 50 µl reaction volume.
