Description
SuperHot Master Mix is an optimized ready-to-use PCR mixture of Taq DNA Polymerase, antibodies to Taq DNA Polymerase, PCR buffer, MgCl 2 and dNTPs. 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.
Master Mix provides reproducible amplification results. The performance of the Master Mix in PCR is not changed after repeated cycles of freezing-thawing.
Simple to handle: Just place it into the tube/plate, add primers and a template, adjust the final reaction volume, mix all components and run PCR.
Performance and purity tests
Tested for the absence of endodeoxyribonucleases and exodeoxyribonucleases. The 2x SuperHot Master Mix is tested for 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 SuperHot Master Mix with 1 µg of pUC19 DNA in 50 µl for 4 hours neither at 37°C nor at 70°C.
Associated activities:
Endonuclease and exonuclease activities were not detectable after 2 hours incubation of the mixture with 0.22 mg of EcoR I digested lambda DNA at 72 °C in the presence of 15 – 20 units of enzyme in SuperHot Master Mix.
Storage conditions
-20°C
Protocol for PCR with SuperHot 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. This 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 |
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.
Feedback from a customer (diagnostic kits producer)
“SuperHot Master Mix gives excellent results in Mycoplasma DNA-detection kits. It was selected by us out of 15 enzymes produced by well-known international suppliers. It shows 100% specificity and very high sensitivity.”
