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Product information "TEMPase Hot Start DNA Polymerase"
About TEMPase Hot Start DNA Polymerase
TEMPase Hot Start DNA Polymerase has been designed to avoid spurious amplification and to diminish the formation of non specific priming events during your set up. TEMPase Hot Start DNA Polymerase allows for detection of low copy number targets and for multiplex PCR in a single reaction tube. TEMPase Hot Start DNA Polymerase is available together with different buffers. This one (A2251xx) is supplied with 10X ammonium sulfate buffer plus 10X combination buffer plus separate 25mM MgCl2.
TEMPase Hot Start DNA Polymerase is also available together with 10X ammonium sulfate buffer and separate 25mM MgCl2 (A2211xx).
Features
- Reaction set up at room temperature
- Increased sensitivity
- Increased specificity
- Increased product yield
Description
TEMPase Hot Start DNA Polymerase is a modified form of Ampliqon Taq DNA Polymerase, which is activated by heat treatment.
A chemical moiety is attached to the enzyme at the active site, which renders the enzyme inactive at room temperature. Thus, during set up and the first ramp of thermal cycling, the enzyme is not active and misprimed primers are not extended.
Once the reaction reaches optimal activating temperature, the chemical moiety is cleaved during a heat activation step, releasing the active TEMPase Hot Start DNA Polymerase into the reaction.
Please check also our SuperHot Hot Start Mastermix > with or without tracking dye.
With our high quality dNTPs as Set (M3015.4100 and M3015.0250) > or Mix (M3016.1010) > or our DNA Ladders > and our favourable standard agarose (M3044) > we can offer additional products for your PCR.
Specifications:
5 units/µL.
shipped with 10X combination buffer plus separate 25mM MgCl2.
Enyzme is completely inactive prior to heating at 95°C for 15 min.!
Application:
for multiplex PCR for Real-time PCR for PCR of low amount of DNA (very low copy number)Units:
One unit is defined as the amount of enzyme which will convert 10 nmoles of dNTPs to an acid-insoluble form in 30 min at 72°C under the assay conditions (25 mM TAPS (tris-(hydroxymethyl)-methyl-amino-propanesulfonic acid, sodium salt) pH 9.3 (25°C); 50 mM KCl; 2 mM MgCl2; 1 mM b-mercaptoethanol; and activated calf thymus DNA as substrate.Source:
recombinantClassification:
Documents:
ManualsGeneral Data 4
Category List
Accessory Items
Agarose LE is a standard agarose for the separation of DNA in the size range between 100bp and 25kbp. It is suitable for all analytical and preparative electrophoresis of nucleic acids in routine gel electrophoresis. Depending on the concentration of Agarose LE used, the size range of nucleic acid separation will vary between 100bp and 25kbp. The low EEO makes this useful for a broad range of applications: PCR product analysis, restriction enzyme digest analysis, separation of RNA before blotting, etc. This agarose is comparable with, e.g. Agarose BioRagent, low EEO from Sigma or with the Universal-Agarose, peqGOLD from Peqlab. Free Taq DNA Polymerase test sample available! No shipping costs within Germany. Genaxxon offers Agaroses for different purposes. Two melting point options and normal or high resolution separation of DNA fragments are available. Standard normal melting agarose M3044 with standard resolution used in routine DNA electrophoresis for separation of a wide range of DNA fragments from 100bp up to 25kbp. This agarose is available as powder (M3044 Agarose LE >) or as high-quality, multi-purpose tablets (M3054 >) available in blister packaging in 200 or 1,000 tablet quantities. They are compact, pre-weight and therefore easy-to-use. Low melting/gelling temperature agarose recommended for rapid DNA gel extraction with the agarose-digesting enzyme, agarase, as well as for "in-gel" DNA treatment with enzymes or for bacterial transformation with nucleic acids directly after re-melting the gel. High resolution agarose (normal melting or low melting) for high resolution elektrophoresis to differentiate DNA fragments with only 2 bp size difference: M3046 Agarose Tiny > (low melt, high resolution comparable to NuSieve®), or M3047 Agarose Tiny HT > (high gel strength, high resolution comparable to SeaKem®). For the nontoxic staining of nucleic acids in agarose gels the highly sensitive fluorescent dye SafeGel red stain (M3193) or SafeGel green stain (M3191) is recommended.
The better performance of the Genaxxon bioscience 5X B-Enhancer Solution compared to standard enhancers such as form amide, DMSO, TMCA or BSA especially when used with GC rich regions or templates with a high degree of secondary structures depends on its ability to lower the DNA melting temperature and its enhancing effect on the DNA polymerase. In detail, Betaine binds preferentially to AT rich sequences in the major groove, thereby stabilizing AT rich regions of the DNA. Because AT forms 2 hydrogen bonds and GC forms 3, the bonding of AT is less stable than the one of GC. As a consequence of the stabilizing effect of Betaine on AT bonding, the stability of AT bonding and GC bonding is brought close to an equal level. At the same time, Betaine has a sequence independent destabilizing effect on all DNA. Summarized, the Tm of AT rich and GC rich sequences are equalized and the overall Tm is lowered. Furthermore, Betaine aids the processivity of thermostable polymerases and reduces "pauses" in polymerization caused by secondary structure that can induce the polymerase to disassociate from the DNA strand.As Betaine has a decreasing effect on the melting temperature of DNA and primers the denaturation temperatures as well as primer annealing temperatures should be reduced by 1 – 5°C compared to the annealing temperature already used. The optimal annealing temperature should be determined individually for each reaction.
Deoxynucleotide (dNTP) Solution Mix as an equimolar solution of ultrapure, HPLC purified (>99%) dATP, dCTP, dGTP and dTTP for qPCR, RT-PCR, standard PCR, and Klenow reactions. The Genaxxon dNTP solutions are optimized for use in DNA amplification and other related methods. The Genaxxon dNTPs and dNTP-mixes contain no measurable bacterial or human DNA. For long term storage and/or for repeated use we do recommend to aliquot the stock solutions. Mixture of dNTP sodium salts (dATP, dCTP, dGTP und dTTP) Concentration: 10 mM each nucleotide Please have also a look on our broad range of nucleotides especially the dNTP mix withv2 mM dNTP mix , our dNTP set, or modified nucleotides, e.g. Biotin-11-dUTP. You can get additional products for your successful PCR as our Taq DNA Polymerse (M3001), or our proof-reading polymerases Pfu (M3004), Pwo (M3002) and ReproFast (M3003), as well as the ready-to-use RedMastermix (M3029), already including dNTPs.
The GenLadder 1kb (ready-to-use) 250bp - 10kbp-ladder is the ideal DNA size marker for fragment sizes from 250 - 10000bp. All bands light up to the same intensity except the 1000 bp and the 3000 bp band which are twice as bright for better orientation. Each band has been calibrated with respect to base pair size and DNA quantity. Thus, an accurate determination of product size and DNA concentration is possible. No ambiguous bands are visible. The GenLadder 250bp - 10kb can be directly labeled at the 5'-end with radioisotopes using T4 polynucleotide Kinase for visualisation by autoradiography. The DNA is already dissolved in buffer with 6X loading dye ready to use. Fragment sizes and amount of DNA per band: 10000bp ( 28ng/5µL), 8000bp (28ng/5µL), 6000bp (28ng/5µL), 5000bp (28ng/5µL), 4000bp ( 18ng/5µL), 2x3000bp (92ng/5µL), 2500bp (34ng/5µL), 2000bp (34ng/5µL), 1500bp (20ng/5µL), 2x1000bp (92ng/5µL), 750bp (23ng/5µL), 500bp (30ng/5µL), 250bp (45ng/5µL). GenLadder DNA ladders are available in a ready-to-use format (premixed with 6X DNA Loading Dye): M3094 (GenLadder 1kbp extended up to 25 kbp ready-to-use) > and M3328 (GenLadder 1kbp ready-to-use) > or our ready-to-use premix M3072 (GenLadder 50bp) > with 6X Orange DNA Loading Dye. The DNA Gel Loading Dyes can also be ordered separately: M3308 (DNA Gel Loading Dye with Bromophenol and Xylene Cyanol FF) >, M3321 (DNA Gel Loading Dye with Orange G) >.
Agarose Tiny is a low-melting high resolution agarose that is suitable for molecular biology applications and produces gels with better separation properties and greater clarity than standard agarose. It is very transparent even at high gel concentrations (3.0 - 4.5%) and can be used with all buffer systems. The low melting temperature makes Agarose Tiny ideal for preparative DNA and RNA electrophoresis, while the low gelling temperature is ideal for tissue culture cell cloning experiments and viral plaque assays. In addition to the above mentioned properties, Agarose Tiny can also be used for high-resolution (2 bp) gel electrophoresis in the range from 10 to 500 bp. Example: PCR products or small DNA fragments after restriction digestion or in mutation analyses. Features:Like Lonza's SeaPlaque™, Agarose Tiny has a low melting point (<64°C, 3.0% gel) and gel point (<25°C, 3.0% gel) and low DNA-binding capacity. Their use is particularly recommended for preparative RNA and DNA gels. Genaxxon offers Agaroses for different purposes. Two melting point options and normal or high resolution separation of DNA fragments are available. Standard agarose high melting point agarose M3044 with standard resolution used in routine DNA electrophoresis for separation of a wide range of DNA fragments from 50bp up to 20kbp. This agarose is available as powder (M3044 Agarose LE >) or as high-quality, multi-purpose tablets (M3054 >) available in blister packaging in 200 or 1,000 tablet quantities. These are compact, pre-weight and therefore easy-to-use. Low melting/gelling temperature agarose recommended for rapid DNA gel extraction with the agarose-digesting enzyme, Agarase, as well as for “in-gel’ DNA treatment with enzymes or for bacterial transformation with nucleic acids directly after re-melting the gel. High resolution agarose (normal melting or low melting) for high resolution elektrophoresis to differentiate DNA fragments with only 2 bp size difference: M3046 Agarose Tiny > (low melt, high resolution comparable to NuSieve®), or M3047 Agarose Tiny HT > (high gel strength, high resolution comparable to SeaKem®). For the nontoxic staining of nucleic acids in agarose gels the highly sensitive fluorescent dye SafeGel red stain (M3193) or SafeGel green stain (M3191) is recommended.Recommended for the following applications:- RFLP (Restriction Fragment Length Polymorphism - AMPFLP (Amplified Fragment Length Polymorphism) - HVR (Hyper-Variable Regionen) - VNTR (Variable Numbers of Tandem Repeats) - STR (Short Tandem Repeats) - Tri und Tetra Nukleotid repeats
Highly pure, HPLC purified (>99%) dNTPs packaged as 4 separate 100mM solutions of dATP, dCTP, dGTP and dTTP for qPCR, RT-PCR, standard PCR, and Klenow reactions. Genaxxon’s dNTP solutions have been optimized for use in DNA amplification and other related methods. Genaxxon dNTPs contain no measurable bacterial or human DNA. For long term storage and/or for repeated use, our recommendation is to aliquot the stock solutions. Solutions of dNTP sodium salts (dATP, dCTP, dGTP und dTTP) Concentration: 100 mM each nucleotide Please have also a look on our broad range of nucleotides especially the dNTP mixes with 2 mM or 10 mM or the modified nucleotides, e.g. Biotin-11-dUTP. You can get additional products for your successful PCR as our Taq DNA Polymerse (M3001), or our proof-reading polymerases Pfu (M3004), Pwo (M3002) and ReproFast (M3003) as well as the ready-to-use RedMastermix (M3029), already including dNTPs.
Our DNA marker GenLadder 100 bp Plus with loading dye is an ideal tool to determine the size of double stranded DNA in the range of 100 - 3000 bp. The additional bands at 1.5kbp and 3.0kbp allow to assign additional DNA fragment sizes in the range up to 3000 bp without using an additional 1kbp DNA marker. The Gene Ladder 100 bp Plus consists of 12 fragments in sizes between 100 - 1000bp spaced 100bp apart. In addition, there are further bands at 1500bp and 3000bp. The 500bp and 1500bp fragment show stronger intensity to allow easier identification. All fragments are 'blunt-ended'. The GenLadder can be directly labeled at the 5'-end with radioisotopes using T4 polynucleotide Kinase for visualisation by autoradiography. Fragment sizes (in base pairs): 3000 bp (40ng/5µL), 2x 1500 bp (70ng/5µL), 1000 bp (50ng/5µL), 900 bp (40ng/5µL), 800 bp (40ng/5µL), 700 bp (30ng/5µL), 600 bp (30ng/5µL), 2x 500 bp (90ng/5µL), 400 bp (40ng/5µL), 300 bp (30ng/5µL), 200 bp (40ng/5µL), 100 bp (40ng/5µL). The DNA marker is already dissolved in buffer containing orange G and Xylene cyanol FF as tracking dyes. The total concentration of the DNA-Marker is 0.1µg/µL (108µg/mL). The recommended amount of DNA marker per lane is 0,5μg (5µL). GenLadder DNA ladders are available in a ready-to-use format (premixed with 6X DNA Loading Dye): M3094 (GenLadder 1kbp extended up to 25 kbp ready-to-use) > and M3328 (GenLadder 1kbp ready-to-use) > or our ready-to-use premix M3072 (GenLadder 50bp) > with 6X Orange DNA Loading Dye. The DNA Gel Loading Dyes can also be ordered separately: M3308 (DNA Gel Loading Dye with Bromophenol and Xylene Cyanol FF) >, M3321 (DNA Gel Loading Dye with Orange G) >.
ROX is used as passive reference dye to compensate for non-PCR related well to well variations in the fluorescence. Variations in fluorescence can occur e.g. due to well to well variation in light intensity, which depends on the optical construction of the PCR machine or due to pipetting variations. The ROX fluorescence does not change during the course of the PCR reaction but provide a stable baseline to which samples are normalized. The excitation and emission of the reference dye are 584nm and 612nm, respectively.The ROX concentration to be used depends on the Real Time PCR instrument or, more precisely, on the filter available. On earlier Real Time PCR machines there was no filter matching precisely to the ROX fluorescence. Therefore, high ROX concentrations are needed with these machines. Some newer machines do not need any internal standard to correct for light intensity. Nevertheless, an internal standard might still be useful to correct for pipetting variations.This dye has been qualified for use on instruments like the ABI PRISM® 7700, One-Step System, RotorGene and many others.Please consult the user manual of the real time PCR instrument with regard to appropriate levels of ROX. Ready to use qPCR Master Mixes with High ROX (500nM) >, Low ROX (50nM) > or without ROX > are also available from Genaxxon.
Accessory Items
Taq DNA Hot Start Polymerase with Aptamer technology. The specific aptamer inhibits DNA polymerase activity below temperatures of 50°C to 55°C, thus enabling hot start PCR. At higher temperatures, the aptamer falls off the active site of Taq polymerase. The Hot Start Taq Polymerase can be used like a normal Taq without having to lengthen the first denaturation step.The binding of aptamer and polymerase prevents the formation of primer-dimer constructs and other artefacts revealed by non-specific primer binding. Check also our SuperHot Mastermix with or without dye. With our high quality dNTPs as Set or Mix or our DNA Ladders and our favourable standard agarose we can offer additional products for your PCR.
GreenMasterMix without ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X Mastermix is ready-to-use and contains optimised amounts of all ingredients. Advantages of the chemically modified SuperHot Taq polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR master mix without ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: BioRad CFX96 Touch™, CFX384 Touch™, CFX Connect™, DNA Engine Opticon® 2, Chromo4™, iCycler iQ™ and My iQ™ , Roche LightCycler® 480, LightCycler® 1536, LightCycler® Nano, LightCycler® 96 and QuantStudio™ instruments, Thermo Scientific™ PikoReal™, Cepheid SmartCycler®, Bio Molecular Systems Mic qPCR cycler, Qiagen Rotor Gene Q, Rotor Gene 6000, MyGo Mini and MyGo Pro. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
ProbeMasterMix without ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X Mastermix is ready-to-use and contains optimised amounts of all ingredients. Multiplex PCR: Applications at Genaxxon and at customers site have shown that the qPCR Probe Mastermix can be used for the simultaneous detection of up to four DNA targets in the same PCR reaction. For further details please refer to the product manual or contact us: info@genaxxon.com. Advantages of the chemically modified SuperHot Taq polymerase used in the Genaxxon ProbeMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Probe qPCR master mix without ROX contains all the necessary components in an optimized composition to carry out quantitative PCR. chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: BioRad CFX96 Touch™, CFX384 Touch™, CFX Connect™, DNA Engine Opticon® 2, Chromo4™, iCycler iQ™ and My iQ™ , Roche LightCycler® 480, LightCycler® 1536, LightCycler® Nano, LightCycler® 96 and QuantStudio™ instruments, Thermo Scientific™ PikoReal™, Cepheid SmartCycler®, Bio Molecular Systems Mic qPCR cycler, Qiagen Rotor Gene Q, Rotor Gene 6000, MyGo Mini and MyGo Pro. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
Highly specific Hotstart master mix for routine and diagnostic PCR and qPCR up to 4kb: ready-to-use master mix with chemical modified Taq DNA polymerase. This Hotstart Master Mix for qPCR is a premixed, ready-to-use solution containing chemically modified Taq DNA Polymerase, dNTPs, MgCl2 and reaction buffers at optimal concentrations for efficient amplification of DNA templates by PCR and qPCR (one tube, one pipetting step). Just add your primers and template DNA. The Hotstart PCR Master Mix has been optimized for use in routine and diagnostic Hotstart PCR and qPCR for amplifying DNA template in the range of 0.2-4kb. The special formulation of our Hotstart Master Mix guarantees you reproducible results even after repeated freezing/thawing steps leading to good yields and sensitivity. The Hotstart Master Mix is efficient (no left over of reagents), scalable from 10µL to 50µL and stable for 12 months. This Hotstart PCR master mix is also available as SuperHot Mastermix Blue > enabling visualisation of each pipetting step. With our high quality dNTPs as Set (M3015.4100 and M3015.0250) > or Mix (M3016.1010) > or our DNA Ladders > and favourable standard agarose (M3044) > we can offer additional products for your PCR.
GreenMastermix High ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X PCR master mix is ready-to-use and contains optimised amounts of all ingredients. Advantages of the chemically modified SuperHot Taq polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR master mix high ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye 500nM ROX optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900 HT, Eppendorf Realplex4, StepOne™ and StepOnePlus™. Other realtime PCR master mixes from Genaxxon are: M3023 - GreenMastermix No ROX >, M3045 - ProbeMastermix No ROX >, M3011 - GreenMastermix Low ROX >, M3031 - ProbeMastermix Low ROX >, M3052 - GreenMastermix High ROX >, M3010 - ProbeMastermix High ROX >.