[TP5000] ExcelTaq™ Hot Start II DNA Polymerase (5 U/μl, 500 U)
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The ExcelTaq™ Hot Start II DNA Polymerase is a mixture of an aptamer-based inhibitor and a recombinant thermo-stable Taq DNA polymerase designed for preventing or minimizing non-specific DNA amplification in PCR reaction. The inactivation of polymerase is achieved by a reversible binding of the aptamer to the polymerase at temperatures below 45°C. The aptamer inhibitor releases polymerase during normal PCR cycling. The aptamer-based inhibition omits the time-consuming initial activation step required by chemically modified or antibody-based hot start polymerases.
The high specificity and sensitivity of ExcelTaq™ Hot Start II DNA Polymerase allows sensitive detection from limited amount of DNA templates, such as 1 pg of cDNA or 1 fg of plasmid DNA. With a high DNA synthesis rate and high thermo-stability, the ExcelTaq™ Hot Start II DNA Polymerase allows reactions to be set up at room temperature and is suitable for common and specialized PCR applications
Detail
Description
The ExcelTaq™ Hot Start II DNA Polymerase is a mixture of an aptamer-based inhibitor and a recombinant thermo-stable Taq DNA polymerase designed for preventing or minimizing non-specific DNA amplification in PCR reaction. The inactivation of polymerase is achieved by a reversible binding of the aptamer to the polymerase at temperatures below 45°C. The aptamer inhibitor releases polymerase during normal PCR cycling. The aptamer-based inhibition omits the time-consuming initial activation step required by chemically modified or antibody-based hot start polymerases.
The high specificity and sensitivity of ExcelTaq™ Hot Start II DNA Polymerase allows sensitive detection from limited amount of DNA templates, such as 1 pg of cDNA or 1 fg of plasmid DNA. With a high DNA synthesis rate and high thermo-stability, the ExcelTaq™ Hot Start II DNA Polymerase allows reactions to be set up at room temperature and is suitable for common and specialized PCR applications
Features
Aptamer-based hot start PCR
Reversible enzyme inactivation
Omits extra enzyme activation step
Convenient for room temperature PCR set-up
High yield and specificity of target amplicons
Wide range of amplicon length (up to 10 kb)
High sensitivity (as low as 1 fg of plasmid)
Applications
High specificity PCR
Generation of PCR products for TA cloning
Routine PCR, multiplex PCR, colony PCR, and RT-PCR
Storage
-20°C for 24 months
Other Products
RevTaq RT-PCR DNA polymerase
Product Info
Document
Product Info
RevTaq RT-PCR DNA polymerase is an engineered, extremely thermostable reverse transcriptase and combined DNA polymerase, obtained through directed, artificial evolution.
RevTaq RT-PCR DNA polymerase can be used similarly to Tth DNA polymerase also for reverse transcription, but does not require the addition of Mn2+ for RT-PCR, which simplifies assay setup and possible sample processing.
– half-life at 95°C of >40 min.
– fast start function due to a hotstart aptamer formulation which prevents unspecific amplification at lower temperatures (<57°C)
– facilitates “zero-step” RT-PCRs directly from RNA templates (without an isothermal reverse transcription step), as reverse transcription takes place simultaneously with DNA amplification during the cycled PCR elongation step.
– allows reverse transcription reactions at high temperatures, thus minimizing the problems encountered with strong secondary structures in RNA that melt at elevated temperatures.
– RevTaq RT-PCR DNA polymerase is the pure, reverse transcription active enzyme ingredient of our Volcano3G® RT-PCR Master Mixes
Please note: Due to the thermostable nature of the enzyme, it is recommended to design very high melting primers and probes (>65°C). See more details in FAQ.
For research use and further manufacturing.
In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.
Document
RevTaq RT-PCR DNA polymerase is an engineered, extremely thermostable reverse transcriptase and combined DNA polymerase, obtained through directed, artificial evolution.
RevTaq RT-PCR DNA polymerase can be used similarly to Tth DNA polymerase also for reverse transcription, but does not require the addition of Mn2+ for RT-PCR, which simplifies assay setup and possible sample processing.
Eleven discrete fragments ranging from 50 bp to 1000 bp
Higher intensity reference band at 500 bp
The Norgen PCR Ranger 100 bp DNA Ladder is prepared to ensure quality and batch-to-batch consistency. Our PCR Ranger DNA Ladder contains eleven discrete fragments ranging from 50 bp to 1000 bp with a higher intensity reference band at 500 bp. This Ladder is ideal for assessment of a range of PCR product sizes.
Contents 1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).
PCR Ranger 100 bp DNA Ladder (Cat# 11300) – 100 loads
Ladder Properties: • Eleven discrete bands, ranging from 50 bp to 1,000 bp • Higher intensity band at 500 bp for easy reference
Fragment
Size (bp)
Mass (ng)
1
1000
91
2
900
66
3
800
61
4
700
51
5
600
43
6
500
70
7
400
28
8
300
23
9
200
30
10
100
22
11
50
15
Recommended Use: Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.
Storage: Stable at room temperature. For longer term storage, -20°C is recommended.
This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.
All-in-one solution for inclusion body protein isolation and purification
Fast and convenient spin column protocol
Complete kit with Cell Lysis Reagent, Inclusion Body Solubilization Reagent, buffers and spin columns to purify proteins
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
These kits provide everything required to isolate and purify inclusion body proteins from induced bacterial cultures. First a proprietary Cell Lysis Reagent is used to selectively lyse the cells and release inclusion bodies in their solid form. Next, inclusion bodies are dissolved and their contents released using the provided IB Solubilization Reagent. Inclusion body proteins are then further purified using spin columns for rapid and convenient buffer exchange and desalting. This kit provides a convenient way to screen recombinants prior to scaling up.
ProteoSpin™ Inclusion Body Protein Isolation Micro Kit
The process is efficient and streamlined and can process up to 12 samples in only 60 minutes. Each spin column is able to recover up to 50 µg of acidic or basic proteins. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.
ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit
The procedure is efficient and streamlined and can process up to 4 samples in approximately 2 hours. Each spin column is able to recover up to 12 mg of acidic or basic proteins from 100 mL of induced bacterial culture. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.
About Inclusion Bodies
Bacteria are widely used for the expression of different proteins. However, 70-80% of the proteins expressed in bacteria by recombinant techniques are typically contained in insoluble inclusion bodies (i.e., protein aggregates). The protein of interest found in these subcellular structures is often inactive, due to incorrect folding. The production rate of recombinant proteins stored in inclusion bodies is invariably higher than those synthesized as soluble proteins. The reason behind this is thought to be the resistance of insoluble proteins to proteolysis by cellular enzymes. In addition, separation of insoluble recombinant proteins in inclusion bodies is considerably easier than that of soluble proteins. These factors have been the major influences favoring scale-up of high-value proteins using bacterial fermentation for example. Procedures for the purification of the expressed proteins from inclusion bodies are often labour-intensive, time-consuming and not cost-effective. This kit provides the essential reagents for cell disruption, inclusion body solubilization and purification using spin column chromatography – all optimized to work together thereby simplifying the process and saving a tremendous amount of time and cost.
The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. Once opened, the solutions should be stored at 4°C when not in use except for the pH Binding Buffers (Acidic and Basic). Some precipitation will occur with 4°C storage. This precipitation should be dissolved with slight heating to room temperature before using.
Component
Cat. 10300 (Micro – 25 preps)
Cat. 17700 (Maxi – 4 preps)
Wash Solution C
30 mL
130 mL
Wash Solution N
30 mL
130 mL
Binding BUffer A
4 mL
20 mL
Binding Buffer N
4 mL
20 mL
Elution Buffer C
8 mL
2 x 30 mL
Protein Neutralizer
4 mL
4 mL
Cell Lysis Reagent
15 mL
110 mL
IB Solubilization Reagent
2 mL
50 mL
Syringes, 1cc, slip tip
25
–
Needles (Bev, 20G x 1 inch)
25
–
Syringes, 10 mL, Luer-Lok™ Tip
–
4
Needles (18G x 1.5 inch)
–
4
Micro Spin Columns
25
–
Maxi Spin Columns (filled with SiC) inserted into 50 mL collection tubes
