• Aptamer-based hot-start functionality
• Immediate hot start without any activation step
• High PCR specificity and sensitivity
• Optimized Master Mix including exceptionally pure ApStarTaq DNA Polymerase and highest quality dNTPs

• Convenient and fast PCR reactions in endpoint and real-time analysis
• Hot-start PCR up to 5 kb
• Amplification of low-copy-number targets
• RT-PCR and TA cloning

biotechrabbit™ ApStarTaq PCR Mix is an optimized aptamer-based hot-start mix and the first-choice for fast PCR reactions. The exceptional quality guarantees highest performance, suitable for standard and fast PCR cycling in both, endpoint and real-time assays. It ensures high product yields with low background and without primer–dimer formation or non-specific priming.

The aptamer binds to Taq DNA Polymerase and inhibits the enzyme activity at temperatures below 45°C. This ensures full hot-start functionality. The enzyme is released during standard PCR cycling conditions. There is no need for separate heating or denaturation steps, allowing fast PCR reactions.

Info: Recommended annealing temperature is 2°C above primer Tm (use gradient PCR to optimize the annealing temperature).

Quality Control

Functional assay

Human genomic DNA was amplified using the ApStarTaq PCR Mix and specific primers to produce a distinct band.

Prevention of PCR contamination

When assembling the amplification reactions, care should be taken to eliminate the possibility of contamination with undesired DNA.

• Use separate clean areas for preparation of samples and reaction mixtures and for cycling.
• Wear fresh gloves. Use sterile tubes and pipette tips with aerosol filters for PCR setup.
• Use only water and reagents that are free of DNA and nucleases.
• With every PCR setup, perform a contamination control reaction that does not include template DNA.

Standard PCR setup

The standard PCR protocol using biotechrabbit reaction buffer provides excellent results for most applications. Optimization might be necessary for certain conditions, such as the amplification of long targets, high GC or AT content, strong template secondary structures or insufficient template purity. In such cases, optimization of template purification (see biotechrabbit nucleic acid purification kits), primer design and annealing temperature is recommended.

The best conditions for each primer-template can be optimized with the following:

• Choosing the optimal quantities of template and primers
• Optimizing cycling conditions

Basic Protocol

• The Master Mix is designed to be used without any optimization as it has all necessary reaction components in optimal amounts for successful PCR.
• Thaw on ice and mix all reagents well.
• Keep all reagents and reactions on ice.
• Pipet the master mix into thin-walled 0.2 ml PCR tubes.
• Add template and primers separately if they are not used in all reactions.

• Mix and centrifuge briefly to collect the liquid in the bottom of the tube.
• Place in the PCR cycler.

Cycling Program

• Add loading dye solution (see DNA Loading Dye, 6×, cat. no. BR0800301) to the reactions to analyze PCR products on a gel or store them at −20°C.