H-SPlus-Taq DNA Polymerase

Hot Start Polymerase with ultra short inactivation time, free of MAB´s. The high concentrated versions are designed for Lyophilization

Cat.-no	Description	Amount	Price €	Shop
S400	H-SPlus Taq DNA Pol. (5 U/µl)	200 units	59.00	add
S410	H-SPlus Taq DNA Pol. (5 U/µl)	1000 units	219.00	add
S410HC25	H-SPlus Taq DNA Pol. HC (25 U/µl)	2000 units *	538.00	contact us
S410HC50	H-SPlus Taq DNA Pol. HC (50 U/µl)	2000 units *	538.00	contact us

* Setting up fee of 450 Euro per preparation/order. Please contact us for details and initial steps.

Hotstart Polymerase H-SPlus: Datasheet
H-SPlus Taq DNA Polymerase: Deutsche Beschreibung

Features:
- Maximo H-SPlus Taq DNA Polymerase is “inactive” at room-temperature
- When the reaction temperature is greater than 70 °C the Polymerase “jumps” into action
- It is neither blocked by antibodies nor by chemically modification -> ultra-short activation time
- For “high-yield” Hot-Start PCR results
- Multiplex PCR
- PCR setting up at room-temperature
- As sensitive Polymerase for PCR Diagnostics research
- ready-to use for lyopholization when using the HC-Versions

Description:
Maximo H-SPlus Taq DNA Polymerase (recombinant from Thermus aquaticus) catalyzes the polymerization of nucleotides into duplex DNA in 5'→3' direction in the presence of magnesium. It also possesses a 5'→3' polymerization-dependent exonuclease replacement activity but lacks a 3'→5' exonuclease activity. The activation of the enzyme starts immediately at 70°C and requires no increased time in heating or denaturation step. Its activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents theextension of nonspecifically annealed primers and primer-dimer formation at low temperatures during PCR setup.

Unit definition:
One unit is defined as the amount of enzyme required to incorporate 10 nmoles of dNTP into acid-insoluble material in 30 min at 72°C.

Concentration: 5 u/µl

Storage buffer: 20 mM Tris-HCl, 100 mM KCl, 0.15 mM EDTA, 1 mM DTT, 0.5% Tween-20, 0.5% Nonidet P40, 50% (v/v) Glycerol, pH 8.1

Reaction buffers :
1.) Reaction buffer (10X)” incomplete” (red cap):160 mM (NH₄)₂SO₄, 670mM TrisHCl pH 8,8, 0,1% Tween-20, enhancers

2.) Reaction buffer (10X) “complete” (yellow cap):160 mM (NH₄)₂SO₄, 670mM TrisHCl pH 8,8, 0,1% Tween-20, enhancers, 25mM MgCl₂3.) separate Tube: 100 mM MgCl₂

Recommended 10 X Reaction Buffer for increased sensitivity (Not provided):
100 mM Tris-HCl (pH 8.8), 500 mM KCl, 0.1 % Tween 20, 15 mM MgCl₂

Quality control
- H-SPlus Taq DNA Polymerase is free from endonucleases and primer contamination, positive PCR performance with several templates of Lambda DNA (<= 12 kb) and human placenta DNA (3kb),

Standard Protocol:

Components

Volume per reaction

10X reaction buffer

5-10 µl

100 mM MgCl₂

optional

dNTP-Mix (40mM)

1.0 µl

Up-stream primer (10 µM stock)

0,5-2.5 µl

Down-stream primer (10µM stock)

0.5-2,5 µl

Template DNA

0.1-15 ng/ml plasmid DNA
1-10 µg/ml genomic DNA

Maximo H-SPlus Taq DNA (5 u/µl)

0.2 - 1.0 µl

Sterile dest. Water (molecular grade)

up to 50 µl total reaction volume

Note:
- vortex all solutions carefully before using
- add the enzyme after Template DNA
- may you have to optimize the MgCl₂ concentration for best result

General Thermo-Cycler protocol:

Step	Time	Temperature
Initial denaturation	1-2	94-95°C
25-30 Cycles: Denaturation Annealing Extension	30 sec 30 sec 0,5-3 min	94-95°C 48-68°C 72°C per 1kb
Final extension	2-3 min	72°C

Note: - In case of low amount of DNA template, additionally cycles may be used

Storage: at -20°C for 24 months, avoid frequent thawing and freezing

Transport: with blue ice or at ambient temperature depending on the destination

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