Hot Start PCR Master Mix (Antibody modified)

Hot Start Taq DNA Polymerase （ Antibody modification ） is a heat-resistant DNA polymerase derived from Thermus aquaticus YT-1. The enzyme has 5’→3′ polymerase activity and double-strand specificity 5 ‘→ 3’exonuclease activity. The enzyme uses antibody modification to block low temperature and room temperature enzyme activity. After thermal activation at 95°C, the antibody 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. The product uses a specially optimized reaction buffer, and HzymesAccu AM Hot Start Taq DNA Polymerase to make a pre-mixed reagent, which maximizes the stability of the enzyme during storage. Only the primer template and water are added before configuring the system, which greatly simplifies The preparation steps.

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 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.