AptaHotTaq is highly suited for qPCR applications. AptaHotTaq minimizes primer-dimer formation, reduces unspecific oligonucleotide priming and therefore, enhances target-specific amplification. AptaHotTaq, which possesses a 5´→ 3´ polymerase activity and a 5´ flap endonuclease activity is a mixture of the thermostable Taq DNA Polymerase with DNA-aptameres inhibiting the enzyme's activity at temperatures up to 50 °C. At temperatures above 50 °C the DNA-aptameres reversibly dissociate from the enzyme. The polymerase is activated during normal cycling conditions, allowing for a convenient assembly of PCR reactions at room-temperature. AptaHotTaq can be used in typical Taq-based cycling protocols. A separate high temperature incubation step to activate the enzyme is not needed but may help to resolve secondary structures of template DNA that may interfere with the initial amplification cycles.

Applications:

•qPCR

•Hot-start PCR

•High throughput applications

Storage Conditions:

-20 °C

Concentration:

5 untis/µl

Recommended PCR Conditions:

Use PCR conditions optimized for Taq DNA Polymerase. In the case of a low amount of DNA template, additional cycles should be used.

Unit Definition:

One unit is defined as the amount of enzyme required to incorporate 10 nmoles of dNTP into acid-insoluble DNA fraction in 30 minutes at 72°C.

Supplied Reaction Buffers:

Complete NH 4 Reaction buffer - Ammonium buffer with 25 mM MgCl 2

Incomplete NH 4 Reaction buffer - Ammonium buffer without MgCl 2

Complete KCl Reaction buffer - Potassium buffer 15 mM MgCl 2

Included MgCl 2 (100 mM)

We recommend Ammonium buffer for increased yield of PCR products and potassium buffer for increased specificity of PCR.

MgCl 2 concentration should be optimized for each particular primer-template combination, meanwhile PCR is effective with 2.5 mM of MgCl 2 in the majority of cases.

• Highly active hotstart Taq

• DNA-aptamer blocked

• Convenient reaction set-up at room temperature

• Activation of enzyme at temperature above 50 °C

• No change or optimization of protocol necessary