Also available at    &     VWR
Reversible 1-min Hot Start Taq DNA Polymerase

Name

Reversible 1-min Hot Start Taq DNA Polymerase

Cat. #

P950
$  59.00
(  100 units)

P950L
$399.00
(1,000 units)

Application

  • Reversible enzyme activation achieve fast and robust PCR.
  • - 1 minute hot start: save 15 minutes !
  • - Proprietary inhibitor: reversible hot start - maximizes specificity
  • - Superior stability: no polymerase activity loss after 1 week RT storage
  • - No freeze-thaw steps: stable at 4oC for 6 months
  • - High sensitivity: PCR can be done with less than 10 copies of DNA templates
Reversible hot start Taq DNA polymerase

Click to see full size image.

This product is for research use only.

Specifications

Polymerase: Reversible 1-min Hot Start
Taq DNA Polymerase
Hot Start: Reversible 1 minute Hot Start
Fidelity: 1 X
GC-Rich PCR Performance: High (with GC-rich buffer)
Amplification Range: 6 kb
Exonuclease Activity: 5´ → 3´
Reaction Speed: 1 kb / 1 min
Reaction Format: Separate Components
Product Overhang: 3'-A

Description and features

Reversible 1-min Hot Start Taq DNA Polymerase is an optimized mixture of superior Taq DNA polymerase and a proprietary inhibitor. The inhibitor reversibly binds to Taq polymerase at temperature below 50°C, and completely inhibits polymerase activity before and after PCR reaction, thus allowing reaction setup at ambient temperature.

1. Fast Hot Start (30 seconds to 1 minute): Initial thermal activation can be achieved by incubating the assembled reaction at 95°C for as short as 30 seconds. Save up to 15 minutes comparing to other Hot Start Taq polymerase, and therefore significantly shortens the reaction time.

2. Exceptional sensitivity: The polymerase can amplify low-abundance template (single-digit copies) in the reaction.

Reversible hot start Taq DNA polymerase

Click to see full size image.

3. Superior stability: The polymerase activity remains the same even after being stored at 25oC for 7 days.

Reversible hot start Taq DNA polymerase

Click to see full size image.

4. Reduced nonspecific amplification: Unlike traditional antibody-based Hot Start Taq polymerase, which loses its “hot start” function once activated, the inhibitor in this product will continue to inhibit the polymerase activity whenever the temperature drops below 50oC, even AFTER the PCR reaction. This unique feature allows researchers to manipulate the PCR product without worrying about nonspecific amplification.

1 minute hot start Taq DNA polymerase

Click to see full size image.

The polymerase has a 5´→3´ DNA polymerase and a 5´→3´ exonuclease activity, and catalyzes the non-template directed addition of an adenine residue to the 3´-end of both strands of DNA molecules, making it suitable for TA cloning. The proprietary polymerase/buffer formulation accommodates extended cycle numbers (45-50 cycles).

Shipping / storage

Ship at 4oC. Store at -20oC for up to 2 years and avoid freeze-thaw cycles. Store at 4oC for up to 6 months.

Quality control

This product is tested for no exogenous nuclease activity; no host DNA contamination tested (by PCR); able to amplify single copy gene from multiple genomes; and no significant enzyme activity decrease after storing at 2 ~ 8oC for 6 months.

Manual (protocol)

  101Bio.com
  Reversible 1-min Hot Start Taq DNA Polymerase

Data sheet

  101Bio.com
  Reversible 1-min Hot Start Taq DNA Polymerase

Components

Components Amount
P950 P950L
Reversible 1-min Hot Start Taq DNA Polymerase 40 µL (100 units) 400 µL (1,000 units)
PCR Buffer 500 µL 4 x 1.5 mL
MgCl2 (100 mM) 100 µL 1 mL
Nuclease-Free Water 1 mL 10 mL
The composition of the 10X PCR buffer is: 100mM Tris-HCl, 500mM KCl, 15mM MgCl2 pH 8.3 @ 25oC

PCR reaction system

Reversible 1-min hot start Taq DNA polymerase

Note: For best results, start with a final concentration of 0.4 μM for Forward and Reverse Primers. For optimization, test the concentration range between 0.15 to 1.0 μM. Magnesium concentration may also be adjusted to achieve best results.

PCR reaction conditions

Reversible 1-min hot start Taq DNA polymerase

Note:

  • To achieve the fastest turnaround, the Hot Start activation can be reduced to 30 seconds.
  • The extension time should be adjusted according to the length of target PCR product size. The amplification speed of this Taq DNA polymerase is approximately 1 kb DNA / 60 seconds.
  • The Annealing temperature should be adjusted based on primer and template composition to achieve the best result.

PCR result examination

After PCR, mix 5 µL PCR product with loading dye and run electrophoresis.

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