Reagents for rapidPCR

If performing a rapidPCR protocoll you would need special polymerases for the rapidPCR. Besides we are offering our optimization kits to adapt the polymerases to your protocol and therefore achieve convenient and reproducible results.

 

Pipette

New designed PCR systems and especially rapidPCR‘s using SpeedCycler2 and AlphaSC®, which allows running times in about 8 minutes, have to be optimized to get the best yield and in addition high specific amplification products. Therefore Analytik Jena offers a matchless tool for the simplest and fastest adaptation of PCR conditions: the SpeedTaq Optimization Set. Each Optimization Set contains a PCR enzyme (standard or
hot start), which are specially optimized for rapidPCR, and a choice of 10 different buffers delivered in pH 8 and pH 9, for the best possible optimization. The ordinary scope of searching the optimal PCR conditions, including the operational effort and the complexity can now be reduced to a minimum without any trouble and loss of quality. Just a few steps have to be performed to get the expected results.

 

Concentration*: 5 U/µl

Enzyme storage buffer: 20 mM Tris-HCl pH 8.0, 100 mM KCl, 1 mM DTT,
0.1 mM EDTA, 0.5% Tween 20, 0.5% Nonidet P40, 50% Glycerol

10x SpeedAmp Buffer Optimization Kit: Buffer 1  –  5 (pH 8 and pH 9), 0.1 ml (each) MgCl2 is already included in the selected buffer, no addition necessary

Store at –20°C, avoid frequent thawing and freezing

Order-No. Amount
845-EZ-1110100 50 units SpeedTaq DNA Polymerase (5U/µl)
10x SpeedAmp Optimization Buffer Kit (10 x 0.1 ml)


*One unit is defined as the amount of enzyme required to catalyze the incorporation of
10 nmol of dNTPs into a polynucleotide fraction in 30 minutes at 70°C.

 

New designed PCR systems and especially rapidPCR‘s using SpeedCycler2 and AlphaSC®, which allows running times in about 8 minutes, have to be optimized to get the best yield and in addition high specific amplification products. Therefore Analytik Jena offers a matchless tool for the simplest and fastest adaptation of PCR conditions: the SpeedTaq Hot Optimization Sets. Each Optimization Set contains a PCR enzyme (standard or hot start), which are specially optimized for rapidPCR, and a choice of 10 different buffers delivered in pH 8 and pH 9, for the best possible optimization. The ordinary scope of searching the optimal PCR conditions, including the operational effort and the complexity can now be reduced to a minimum without any trouble and loss of quality. Just a few steps have to be performed to get the expected results.

 

Concentration*: 5 U/µl

Enzyme storage buffer: 20 mM Tris-HCl pH 8.0, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5% Tween 20, 0.5% Nonidet P40, 50% Glycerol

10 x SpeedAmp Buffer Optimization Kit: Buffer 1  –  5 (pH 8 and pH 9), 0.1 ml (each)  MgCl2 is already included in the selected buffer, no addition necessary

Store at –20°C, avoid frequent thawing and freezing

Order-No. Amount
845-EZ-1120100 50 units SpeedTaq HOT DNA Polymerase (5 U/µl)
10x SpeedAmp Optimization Buffer Kit (10 x 0.1 ml)


*One unit is defined as the amount of enzyme required to catalyze the incorporation of
10 nmol of dNTPs into a polynucleotide fraction in 30 minutes at 70°C.

 

The SpeedTaq DNA Polymerase is specially optimized for extreme fast PCR. Extension times are reduced to a minimum due to a synthesizing speed of 200 bp/sec, what in return dramatically reduces the total reaction time. The SpeedTaq is a mixture of an ultrapure Taq Polymerase with a second thermal stable polymerase. This combination ensures utmost purity, reproducibility and processivity. Thus it is optimal for amplifications running on rapid thermal cyclers, as the SpeedCycler family from Analytik Jena | bio solutions.

 

Concentration*: 5 U/µl

Enzyme storage buffer: 20 mM Tris-HCl pH 8.0, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5% Tween 20, 0.5% Nonidet P40, 50% Glycerol

PCR Buffer: 10x SpeedAmp Buffer (1 ml), buffer type has to be determined in advance using SpeedTaq Optimization Set (Order-No.: 845-EZ-1110100) MgCl2 is already included in the selected buffer, no addition necessary

Store at –20°C, avoid frequent thawing and freezing

Order-No.
 Amount
845-EZ-5000500 500 units


*One unit is defined as the amount of enzyme required to catalyze the incorporation of
10 nmol of dNTPs into a polynucleotide fraction in 30 minutes at 70°C.

 

The SpeedTaq HOT DNA Polymerase is specially optimized for rapidPCR but also provides improved specificity and sensitivity when amplifying low-copy-number targets in complex backgrounds or when prolonged room temperature set up is required. Due to a synthesizing speed of 200 bp/sec, extension times and in return the total reaction time is reduced to a minimum. The polymerase activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents the extension of non-specifically annealed primers and primer-dimer formation  at low temperatures during PCR setup. Thus it is optimal for amplifications running on rapidPCR thermal cyclers, as the SpeedCycler family from Analytik Jena | bio solutions. The enzyme 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. SpeedTaq HOT DNA Polymerase requires no prolonged heating or denaturing step. The polymerase inhibiting ligand is quickly released at the increased temperature of thermal cycling.

 

Concentration*: 5 U/µl

Enzyme storage buffer: 20 mM Tris-HCl pH 8.0, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5% Tween 20, 0.5% Nonidet P40, 50% Glycerol

PCR Buffer: 10x SpeedAmp Buffer (1 ml), buffer type has to be determined in advance using SpeedTaq HOT Optimization Set (Order-No.: 845-EZ-1120100); MgCl2 is already included in the selected buffer, no addition necessary

Store at –20°C, avoid frequent thawing and freezing

Order-No. Amount
845-EZ-7000500 500 units


*One unit is defined as the amount of enzyme required to catalyze the incorporation of
10 nmol of dNTPs into a polynucleotide fraction in 30 minutes at 70°C.

 

RapidPCR allows PCR reactions in about 8 minutes by using the devices of the SpeedCycler family. This poses a challenge for any Taq DNA Polymerase. The SpeedAmp buffers were designed especially to optimize the function of Taq DNA Polymerases for the use of such rapidPCR thermal cyclers. The kit contains a choice of 10 different buffers to obtain the best performance for the reaction. Just select the suitable buffer for the application, which is also available separately.

For best possible optimization, the buffers are delivered with pH 8 and pH 9.
Note: The optimization buffers already contain MgCl2.Each SpeedAmp Optimization Buffer Kit contains 5 different 10x buffers pH 8 and 5 different 10x buffers pH 9.

 

Simple PCR Optimization using SpeedCycler family
The amplification of an E. coli rRNA gene using SpeedAmp Optimization Buffer Kit (buffer
1 – 5, pH 8) shows typical results of the first rapidPCR run. Thereby the fragments ampli-
fied with buffer 1 and 2 are poor in yield. The amplification with buffer 4 and 5 led to an
unspecific primer binding. Consequently the buffer of choice is the buffer No. 3. Additional
lane 1 represents a negative control in comparison to the expected PCR products.


Store at –20°C, avoid frequent thawing and freezing

 

 

Order-No. Amount
845-AK-1100100 10 x 0.10 ml
845-AK-1100250 10 x 0.25 ml
845-AK-1100500 10 x 0.50 ml

 

 
 
Order-No. for 0,5 ml Bestellnummer for 1,0 ml Product/Description
845-AK-1200500
845-AK-1200501		 845-AK-1201000
845-AK-1201001
 Buffer 1, pH 8
Buffer 1, pH 9
845-AK-1300500
845-AK-1300501		 845-AK-1301000
845-AK-1301001
 Buffer 2, pH 8
Buffer 2, pH 9
845-AK-1400500
845-AK-1400501		 845-AK-1401000
845-AK-1401001
 Buffer 3, pH 8
Buffer 3, pH 9
845-AK-1500500
845-AK-1500501		 845-AK-1501000
845-AK-1501001
 Buffer 4, pH 8
Buffer 4, pH 9
845-AK-1600500
845-AK-1600501		 845-AK-1601000
845-AK-1601001		 Buffer 5, pH 8
Buffer 5, pH 9

Further information will follow soon.

 

 

The Speed ready mix is a PCR master mix containing a rapidPCR-optimized amplification
buffer, a rapidPCR-optimized Mg²+  amount and dNTP‘s (dATP, dCTP, dGTP, dTTP).
The master mix offers a simple workflow and faster turnaround. It is suitable for researchers who want to save time and increase sample throughput without compromising the high quality of PCR analyses with the SpeedCycler²  or the AlphaSC. The Speed ready mix can also be used for real-time assays without an Uracil-DNA Glycosylase. The delivered amount of Speed ready mix is sufficient for 100 amplification reactions à 15 µl or 60 amplification reactions à 25 µl.

 

Store at –20°C, avoid frequent thawing and freezing

 

Order-No.
 Amount
845-RM-1000300 For 100 amplification reactions

 

 

The Speed ready mix plus is a PCR master mix containing a rapidPCR-optimized amplification buffer, a rapidPCR-optimized Mg²+ amount and dNTP‘s (dATP, dCTP, dGTP, dTTP). The Speed ready mix plus contains also a DNA Polymerase. The master mix offers a simple workflow and faster turnaround. It is suitable for researchers who want to save time and increase sample throughput without compromising the high quality of PCR analyses with the SpeedCycler²  or the AlphaSC®. The Speed ready mix can also be used for real-time assays without an Uracil-DNA Glycosylase.  The delivered amount of PCR ready mix plus and innuTaq DNA Polymerase is sufficient for 100 amplification reactions à 15 µl or 60 amplification reactions à 25 µl.

 

Store at –20°C, avoid frequent thawing and freezing

 

Order-No.
 Amount
845-RM-1100300 For 100 amplification reactions