Hot Start Taq DNA Polymerase Super Mix (Chemically Modified)
Product Information
| Product name | Cat. No. | Size |
| Hot start Taq DNA | HMD3701S | 100 T |
| polymerase Supermix (chemical modification) | HMD3701M | 500 T |
Product Description
Taq DNA polymerase is a heat-resistant DNA polymerase derived from Thermus aquaticus YT- 1. The enzyme has a polymerase activity of 5’→ 3’and a specificity of 5’→ 3 for the double-strand ‘Exonuclease activity. The enzyme uses a chemical modification group to block the enzyme activity at low temperature and room temperature.
After thermal activation at 95°C, the chemically modified group falls off to release the enzyme activity. Before the amplification system is formulated to PCR amplification, non-specific amplification due to primer mismatch will not occur,
which minimizes the formation of non-specific amplification and primer dimers, and improves the sensitivity and specificity of amplification. This product uses a specially optimized reaction buffer and hot-start Taq DNA.
polymerase (chemically modified) to make a premixed reagent, which maximizes the stability of the enzyme during storage. Only the primer template and water are added before configuring the system. , Greatly simplifies the preparation steps. This product has strong versatility and is suitable for end-point PCR, dye, and probe qPCR.
Product components
| Component | HMD3701S | HMD3701M |
|---|---|---|
| 2 ×Hot start Taq DNA polymerase Supermix (chemical modification) | 1 mL | 5×1 mL |
Transportation and storage methods
≤0℃ for transportation; -20℃ for storage.
Unit definition
Using activated salmon sperm DNA as a template/primer, the activity of ingesting 10 nmol of total nucleotides as acid-insoluble at 74°C in 30 min is defined as 1 activity unit (U).
Quality Control
- Endonuclease residue detection: 4 μg pUC19 DNA is added to this reagent and incubated at 37 ℃ for 4 h, the electrophoresis band of the DNA does not change.
- 5 kb λDNA amplification detection: After 25 cycles of amplification with 5 ng λDNA as a template, agarose gel electrophoresis shows a single specific 5 kb band.
- Exonuclease detection: add 1μM 5′-FAM-labeled oligonucleotide to 20μL reaction system, and incubate at 37°C for 30min without degradation signal.
- Heat inactivation: None detected
- Storage stability: Repeated freezing and thawing of this reagent 50 times, the amplification performance will not decrease.
Reaction system
| Component | volume |
|---|---|
| Template DNA a | optional |
| 10 µM Forward primer | 0.5 μL |
| 10 µM Reverse primer | 0.5 μL |
| 2×Hot start Taq DNA polymerase Supermix (chemical modification) | 10 μL |
| Nuclease-free water | Up to 20 μL |
a:
| DNA | Amount |
| Genome | 1ng – 1μg |
| Plasmid or virus | 1pg – 1 ng |
b: The primer volume is the recommended dosage, which can be optimized according to the actual situation.
Reaction program:
| Step | Temperature | Time | Number of cycles |
|---|---|---|---|
| Predenaturation | 95℃ | 10 min | — |
| denaturation | 95℃ | 15-30s | |
| annealing | 45-68℃ | 15 – 60s | |
| extend | 68℃ | 1 kb/min | |
| Final extension | 68℃ | 5 min | — |
a: The pre-denaturation time for common templates is 1 min, the recommended time for difficult templates is 2-3 min, and the colony PCR pre-denaturation time is recommended to be 5 min.
b: The recommended annealing time is 15-60s. The annealing temperature is generally determined at 45-68℃ according to the Tm of the primer, and it is generally set to be 3-5℃ lower than the Tm of the primer.
The Certificate of Analysis (COA) is a signed document that includes the storage temperature
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