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.
Transportation and storage methods
≤0℃ for transportation; -25～-15℃ for storage.
Unit definition
Using activated salmon sperm DNA as a template/primer, the activity of ingesting 10nmol of total nucleotides as acid insolubles 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
a:
b: The primer volume is the recommended dosage, which can be optimized according to the actual situation.
a: The pre-denaturation time for common templates is 10 min, the recommended time for difficult templates is 15 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) & Material Safety Data Sheet (MSDS) Is A Signed Document That Includes The Storage Temperature,