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Enzymes for Molecular Biology

PCR & DNA Amplification

Taqs like it hot and some like it hot from the start

Most in vitro amplification applications use a thermostable enzyme for the polymerase chain reaction (PCR). The most used PCR enzyme is Taq DNA polymerase. Taq DNA polymerase is derived from Thermus aquaticus, an extremely thermophilic eubacterium which grows at temperatures above 70°C. This means that the optimal enzymatic activity of Taq DNA polymerase is expressed at approximately 72°C. 

Standard PCR or hot-start?

Standard Taq polymerase expresses optimal activity at 72°C but the enzyme becomes active from around 37°C. The low-temperature activity is substantial and has the disadvantage of permitting nonspecific amplification and mis-priming events during the initial lower-temperature phase of a PCR reaction. If your needs include automation of PCR, then consider using a hot-start Taq polymerase.

Hot-start polymerases, unlike standard Taq, are unreactive at ambient temperatures. Hot-start PCR techniques follow the same principles as conventional PCR, but enzymatic activity is suppressed until the first denaturation step has been accomplished. This minimizes nonspecific binding and mis-priming and results in improved specificity. In addition, the technology offers the significant convenience of reaction set up at room temperature making PCR automation possible. In some cases, even sensitivity is improved, because non-specific amplification depletes reaction components required for specific target amplification.

All Taq polymerases are suitable for:

Choose a Taq DNA polymerase based on your requirements for standard PCR or for a hot-start enzyme suitable for automation and high throughput PCR applications.
Enzyme   Characteristics Supplied with

  Hot-start
(x min @ 95°C)		 Amplicon size Extension rate 5’-3’
exonuclease		 PCR buffer Loading buffer
P7250L
Taq-B (Taq)
 Coming soon ✘ No <5 kb
 2–4 kb/min
at 72°C
 ✔ Yes ✘ No ✘ No
P7620L
TaqIT *(Stoffel)
 Coming soon ✘ No <7 kb 2–4 kb/min
at 72°C		 ✘ No* ✔ Yes ✘ No
201203
Taq
   ✘ No <7 kb 2–4 kb/min
at 72°C		 ✔ Yes ✔ Yes + Q‑Solution ✔ Yes
201223
Taq PCR Core Kit
   ✘ No <7 kb
 2–4 kb/min
at 72°C  		✔ Yes  ✔ Yes + dNTPs ✔ Yes
203123
AllTaq PCR Core Kit
   ✔ Yes (2 min.) <9 kb 1 min/kb
at 68°C		 ✔ Yes ✔ Yes + dNTPs ✔ Yes
203203
HotStarTaq		   ✔ Yes (15 min.) <7 kb 2–4 kb/min
at 72°C		 ✔ Yes ✔ Yes + Q‑Solution ✘ No

FAQs about enzymes for PCR and DNA amplification

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