dU-HOT Mastermix
| Catalog # | Pack size | Price, Euro | SHOP |
| 119202 | 100 rcs (2x1,25ml) | 65,00 | Add to cart |
| 119210 | 500 rcs (10x1,25ml) | 320,00 | Add to cart |
PCR-Mastermix information : The Master Mixes are calculated for 50 µl per reaction. Your advantage: if you work with 25 µl reaction volume you can run 200 tests.
Description:
2x dU-HOT Master Mix is an optimized ready-to-use PCR mixture of Taq DNA Polymerase, and antibodies to Taq DNA polymerase, PCR buffer, MgCl2 and dNTPs. dUTP in the Mixture provides the tool for carry-over protection of the sample with UDG (cat#111025) treatment. 2x 
PCR Master Mix contains all components for PCR, except DNA template and primers. The mixture was shown to be effective for Real Time PCR.
2x dU-HoT Master Mix Composition
- Taq DNA Polymerase in reaction buffer: 0.1 units/µl
- Antibodies to Taq DNA polymerase, concentration adjusted for the effective inhibition of polymerase activity at 37oC
- 32 mM (NH4)2SO4
- 130 mM TrisHCl, pH 8.8 at 25oC
- 0,02% Tween-20
- 3 mM MgCl2
- dNTPs (dATP, dCTP, dGTP, dTTP): 0.4 mM of each
- dUTP - 0,6 mM
Pack size:
2x 1,25 ml - sufficient for 100 HotStart PCR reactions in 50 µl reaction volume
10x 1,2 5ml - sufficient for 500 HotStart PCR reactions in 50 µl reaction volume.
Supplied with the enzyme: 1 ml tube MgCl2 (100 mM)
Performance and purity tests:
Tested for the absence of endodeoxyribonucleases and exodeoxyribonucleases. The 2x dU-HOT Master Mix is tested in the amplification of a single-copy gene of mouse genomic DNA.
Endodeoxyribonuclease Assay:
No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 25 µl of 2x dU-HOT Master Mix with 1 µg of pUC19 DNA in 50 µl for 4 hours neither at 37°C nor at 70oC.
Protocol for PCR with dU-HOT Master Mix PCR Master Mix
Due to the inhibition of polymerase activity at room temperature by Anti Taq DNA polymerase antibodies all reactions may be settled-up at room temperature, it will not result in increase of unspecific product or primer-dimers formation.
Add in a thin walled PCR tube:
50 µl reaction volume | 25 µl reaction volume | |||
| component | volume | final conc. | volume | final conc. |
| 2x PCR Master Mix | 25 µl | 1x | 12.5 µl | 1x |
| Forward Primer | variable | 0.1-1 µl | variable | 0.1-1 µl |
| Reverse Primer | variable | 0.1-1 µl | variable | 0.1-1 µl |
| Template DNA | variable | 10 pg-1µg | variable | 10 pg-1 µg |
| 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 a PCR program.
