- 2x Hot Start Taq Mix: 1 ml
- Nuclease-Free Water: 1 ml
Hot Start 2X Mix is a pre-mixed, ready-to-use solution containing Hot Start Taq DNA Polymerase, dNTPs, Mg2+ and Reaction Buffer at optimal concentrations for efficient amplification of DNA templates by PCR. To prepare the final PCR, only primers and template DNA should be added. Hot Start Mix contributes to highly reproducible PCR by reducing the risk of pipetting errors, miscalculation and contamination. It also contributes to higher specificity by optimizing the system, reducing primer-dimer rate. The mix has dramatic increased the sensitivity by adding enhancer.
Hot Start Taq DNA Polymerase is a thermostable recombinant DNA polymerase derived from thermophilic bacterium Thermus aquaticus .Its molecular weight is 94 kDa. Hot Start Taq DNA Polymerase can amplify DNA target up to 5 kb. The elongation velocity is 0.9~1.2kb/min. It has 5' to 3' polymerase activity but lacks of 3' to 5' exoneclease activity, which results in a 3'-dA overhangs PCR product. All components of the Hot Start Mix are at optimal concentration for efficient amplification, which contributes to highly specific incorporation of primer and template.
Convenient: Medico recombinant Taq DNA Polymerase in a ready-to-use mix
Fast: saves time due to reduced number of pipetting steps
Reproducible: lower contamination and pipetting error risk
Compatible with TA cloning: generates PCR products with 3'-dA overhangs
High throughput PCR
Routine PCR with high reproducibility
Generation of PCR products for TA cloning
The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases is confirmed by appropriate quality tests. Functionally tested in amplification of a single-copy gene from human genomic DNA.
One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nM of dNTPs into an acid-insoluble form in 30 minutes at 70°C using hering sperm DNA as substrate.
Composition of the Hot Start Mix:
0.3U/ul Hot Start Taq DNA polymerase
2xHot Start PCR Buffer
0.4mM dNTPs, 3.2mM MgSO4
0.02% bromophenol blue.
Store at -20°C
Repeated freeze-thaw cycles do not reduce the activityof the reactions.
Hot Start Taq Mix (2X)
Ships in: 5 Business Days
Protocol for PCR
All solutions should be thawed on ice, gently vortexed and briefly centrifuged.
Add in a thin walled PCR tube on ice:
For a total 50μl reaction volume
Component of sample Volume Final concentration Hot Start Taq Mix (2X) 25 μl 1X Forward Primer variable 0.1 - 1 μM Reverse Primer variable 0.1 - 1 μM Template DNA variable 10 pg - 1 μg Water, nuclease-free to 50 μl -
For a total 25 μl reaction volume
Component of sample Volume Final concentration Hot Start Taq Mix (2X) 12.5 μl 1X Forward Primer variable 0.1 - 1 μM Reverse Primer variable 0.1 - 1 μM Template DNA variable 10 pg - 1 μg Water, nuclease-free 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. This step may be omitted if the thermal cycler is equipped with a heated lid.
·Place samples in a thermocycler and start the program.