Application

1. Hot start PCR amplification of all various templates or tasks ;
2. designed for amplification of difficult templates, such as GC-rich fragments and microsatellites;
3. primer extension of SNP markers;
4. amplification of genomic DNA targets up to 10 kb with high fidelity, specificity, and sensitivity;
5. high through-put Hot Start PCR with high specificity, sensitivity, and yield;
6. routine diagnostic Hot Start PCR requiring high reproducibility.

DESCRIPTION
HotStart PCR Ready Mix is a 2 x concentrated reagent mix for Hot Start PCR comprising two cold sensitive mutant Taq polymerases, 2 x PCR-buffer with 5 mM MgCl2, 400µM of each dNTP and stabilizers. Unlike monoclonal antibody based hot start Taq or chemical modified hot start Taq, cold sensitive mutant Taq will retain the hot start ability throughout the whole amplification cycles.

Cold sensitive mutant Taq polymerases are designed for "Hot Start" PCR due to their suppressed activity at low temperature. It offers excellent specificity and two-fold higher fidelity than wild-type Taq. It is designed for PCR with difficult templates such as GC-rich fragments and microsatellites. Cold sensitive mutant Taq polymerases are particularly well suited to primer extension of Single Nucleotide Polymorphism (SNP) markers.

Cold sensitive mutant Taq polymerases maintain excellent specificity and minimal background even in conditions designed for high yield (high Mg²⁺ /primer concentrations). In fact, even on genomic templates, the enzyme can be used with MgCl²⁺ concentrations as high as 10 mM.

Cold sensitive mutant Taq polymerases are capable of extending through difficult regions, e.g. regions, which include inverted tandem repeats and those with high amounts of secondary structure.

Cold sensitive mutant Taq polymerases work in a totally unique way, involving improved nucleotide selection at the active site, and a much lower rate of mis-match extension, meaning that only perfectly aligned primers will be extended. As a result, the enzyme can give even higher specificity than hot-start (manual or automatic) techniques without the need for inconvenient pre-incubation steps.

COMPONENT

Cat# DP-008-1025, 250 units Taq (100 U per ml ready mix):

2 x HotStart PCR ready mix 2 x 1.25 ml

ddH₂O 2 x 1.5 ml

Cat# DP-008-1100, 1000 units Taq (100 U per ml ready mix):

2 x HotStart PCR ready mix 8 x 1.25 ml

ddH₂O 8 x 1.5 ml

STORAGE TEMPERATURE

Recommended to keep at 2-8°C for immediate use (12 month) and -20°C for long term storage.

QUALITY CONTROL

Activity, non-specific endonucleases, nickases, and exonucleases.