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End-Point PCR & RT
End-Point PCR Kits
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End-Point PCR Kit


TAQXpedite™ PCR System (FAST End-Point)

Applications
  • Fast PCR with a standard PCR thermocycler.
  • Fast PCR of templates with up to 80% GC content, amplicons longer than 30 kb, and multiplex PCR.
Fast PCR can be achieved using fast PCR instrumentation or by using enzyme/reagent combinations that improve PCR efficiency. With a standard thermocycler, the TAQXpedite™ PCR system achieves fast PCR results with total reaction time as little as 16 minutes for a 500-bp amplicon (Table 1, Fig. 1). The TAQXpedite™ kit can be used with a either a specialized fast PCR thermocycler or a standard thermocycler.

The TAQXpedite PCR System (FAST End-Point) contains a unique blend of thermostable DNA polymerases that can be used for fast PCR, a carefully optimized 2X Universal MasterMix and a Difficult/Long MasterMix with all four dNTPs, and an optimized MgCl2 concentration. The MasterMix also contains Epicentre's patented PCR Enhancer with betaine,* which substantially improves the yield, efficiency, and specificity of amplification of many target sequences, especially those containing a high GC content or secondary structure.1-5 The TAQXpedite PCR System's Universal Master-Mix is designed for routine and/or high-throughput fast PCR applications (Fig. 1), while the Difficult/Long MasterMix is designed for use with troublesome, non-routine, or long applications (Fig. 2). All reaction components are included in the kit except for template and primers.

Benefits
  • Increased sample throughput.
  • High fidelity due to proofreading capabilities.
  • Two convenient MasterMixes for universal and difficult/long amplifications.

g_taqxpeditepcr_fastpcr.jpg
 Figure 1. Fast PCR of a 539-bp amplicon from lambda DNA. Different starting concentrations of template were used in a 16-minute reaction with the TAQXpedite™ kit. Lanes M, 100-bp ladder; lane 1, 100 pg; lane 2, 10 pg; lane 3, 1 pg.

g_taqxpeditepcr_systemcomparison.jpg
 Figure 2. Comparison of the TAQXpedite™ PCR System with competitive kits for amplification of a 4-kb fragment. Gel image depicts results starting with 1 ng lambda DNA as the template for PCR. Lane M, DNA marker; lane 1, TAQXpedite PCR System; lane 2, Competitor Q's Fast Kit; lane 3, Competitor A's Fast PCR MasterMix.

Table 1. Total amplification times for TAQXpedite™ and standard PCR protocols.
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References
  1. Schanke, J.T. and Grunenwald, H.L. (1997) Epicentre Forum 4(1), 2.
  2. Grunenwald, H.L. and Schanke, J.T. (1997) Epicentre Forum 4(1), 4.
  3. Mytelka, D.S. and Chamberlin, M.J. (1996) Nucleic Acids Res. 24, 2774.
  4. Henke, W. et al. (1997) Nucleic Acids Res. 25, 3957.
  5. Weissensteiner, T. and Lanchbury, J.S. (1996) BioTechniques 21, 1102.
TAQXpedite™ PCR System. Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser's own internal research. No other patent rights (such as 5´ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

*Covered by issued and/or pending patents.


FailSafe™ PCR System

Applications
  • Simplified PCR optimization.
  • PCR of difficult or high-GC templates (Fig. 1).
  • Multiplex PCR (Fig. 2).
  • PCR amplifications up to 20 kb for cloning.
The FailSafe™ PCR System provides dependable, consistent high-fidelity PCR results for every DNA template, regardless of its source or sequence. The FailSafe PCR System will faithfully amplify your template every time.
Using Epicentre's patented PCR Enhancement Technology, the FailSafe PCR PreMix Selection Kit has replaced the need for cumbersome adjustments of reaction components to determine the optimal conditions for PCR of each template and primer pair. The 12 FailSafe PCR PreMixes in this kit cover a meticulously determined matrix of PCR conditions that give optimal PCR results for different sequences. Each PreMix represents a unique PCR condition, with everything needed for successful PCR. Combine the template and primers with the FailSafe Enzyme Mix, add one volume of this cocktail to one volume of each of the 12 FailSafe PCR 2X PreMixes, and cycle. Epicentre guarantees that at least one FailSafe PCR PreMix will provide conditions that are optimal for the template and primers tested.
For reliable and consistent PCR results for all subsequent amplifications using the same primers, simply order the FailSafe PCR System with PreMix Choice, and specify the same FailSafe PCR PreMix that gave optimal PCR results using the FailSafe PreMix Selection Kit.

Benefits
  • Successful PCR, the first time and every time.
  • High PCR accuracy, using a unique PCR Enzyme Mix with a 3-fold lower error rate than Taq DNA polymerase.
  • Extremely high sensitivity and specificity using the PCR Enhancer Technology.
  • FailSafe PCR PreMixes contain all components needed for successful PCR.
  • Generates PCR products suitable for both TA cloning and blunt-end cloning.

i_failsafe_neverfail.gif

"Epicentre products are great, as for FailSafe™ PCR, we use it all the time."
Prof. Sir Martin Evans, FRS
Nobel Laureate, Cardiff University, Wales, UK

g_failsafe_humanfragilexgene.jpg
 Figure 1. Amplification of an 80%-85% GC-rich region of the human fragile X gene. PCR was performed using the FailSafe™ PCR System, schematically depicted above. Lanes A-L show the amplification products resulting from PCR using the 12 FailSafe PCR PreMixes. Lane M, molecular weight marker. In this experiment, optimal amplification was obtained with FailSafe PCR PreMix J. The size of the expected amplicon is indicated by an arrow.

 Figure 2. Multiplex PCR of the human CFTR gene. The FailSafe™ PCR System amplified all five exons of the CFTR gene from as little as 1 ng of human genomic DNA. Lane M, DNA marker; lane 1, negative control; lanes 2-5, multplex PCR from 1, 10, 50, and 100 ng, respectively, of human genomic DNA.
g_failsafe_multiplexpcr.jpg

 
fragile_x.jpg
 Figure 3. FailSafe™ PCR of 85+%-GC human fragile X gene. Fragile X gene was amplified from as little as 1 ng of total human genomic DNA.

gel-sensitivity-cropped.jpg 
 Figure 4. Detection of the cII gene in one copy of the phage lambda DNA genome.


 
g_failsafepcr_fig5.jpg
 Figure 5. FailSafe™ PCR of a range of different sequences and sequence sizes in phage, human and bacterial DNA. PCR products shown are up to 20 kb for lambda DNA, up to 21.5 kb for human DNA, and up to 18 kb for E. coli DNA.



FailSafe™ PCR Premix Selection Kits: Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser's own internal research. No other patent rights (such as 5´ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.


FailSafe™ PCR PreMixes (reorder)

This listing is only for reordering of individual FailSafe™ PCR 2X PreMixes.
We recommend first optimizing PCR conditions using the FailSafe™ PCR System.


MasterAmp™ Extra-Long PCR Kit

Applications
  • High-fidelity PCR amplification of sequences up to ~40 kb.
The MasterAmp™ Extra-Long PCR Kit is a complete system for successful and accurate one-step amplification of DNA sequences up to ~40 kb. The kit also contains nine MasterAmp Extra-Long PCR 2X PreMixes with dNTPs, buffer, and varying amounts of MgCl2 and the MasterAmp PCR Enhancer with betaine.* Perform Step 1 (Fig. 1) with each new template and primer set. One PreMix will amplify the long template and give consistent results in all subsequent amplifications (Step 2, Fig. 1).

Benefits
  • Fast optimization; Extra-Long PCR 2X PreMixes with MasterAmp PCR Enhancer, as well as a robust and high-fidelity DNA polymerase mix, allow optimized extra-long PCR (Fig. 2).

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g_masterampxl_lambdadna.jpg

 
 A. Amplification of a 20-kb region of lambda DNA
using MasterAmp™ Extra-Long PCR 2X PreMixes (1-9). MasterAmp™ Extra-Long PCR PreMix 4 produced optimal results.

             
g_masterampxl_subsequentpcr.jpg
B. Subsequent PCR
of a 20-kb lambda DNA region with the same primer pair and MasterAmp™ PreMix 4 gave consistent results, using the optimal PreMix determined in Step 1.
Figure 1. Two steps to reliable and consistent PCR results.

g_masterampxl_xlamplification.jpg
 Figure 2. Extra-long amplification of lambda DNA using the MasterAmp™ Extra-Long PCR Kit. PCR conditions were optimized and MasterAmp Premix 4 was selected for PCR of 1 ng of lambda DNA in each sample. Results were analyzed on a 0.5% agarose gel. Amplicon sizes were 30 kb (lane 1), 35 kb (lane 2) and 40 kb (lane 3). Lane M, 5-Kb DNA ladder.

*Covered by issued and/or pending patents.
MasterAmp™ Extra-Long PCR Kit; Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser"s own internal research. No other patent rights (such as 5´ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.


MasterAmp™ PCR Optimization Kits

Applications
  • Simplified PCR optimization with a thermostable DNA polymerase of choice.
  • PCR of difficult or high-GC templates.
  • PCR amplifications up to 20 kb.
  • Multiplex PCR.
For rapid, dependable optimization of PCR, we encourage you to try the FailSafe™ PCR System to obtain the best PCR results with any template up to ~20 kb. However, if you prefer to use another enzyme, Epicentre also has two MasterAmp™ PCR Optimization Kits that will help you to obtain the best possible PCR results with thermostable enzymes frequently used in PCR, such as Taq, Pfu, Tth, Tfl, and AmpliTherm™ DNA polymerases. These MasterAmp PCR Optimization Kits contain a series of 12 MasterAmp PCR 2X PreMixes that have been meticulously tested and represent a complete range of PCR conditions frequently encountered. Each MasterAmp 2X PreMix contains all components needed for PCR except for the primers, the template, and the PCR enzyme.

First, perform PCR with your template, primers, and the 12 MasterAmp PreMixes. Select the PreMix that gives optimal results. Then, use the same MasterAmp PCR PreMix to obtain reliable, consistent PCR of the same sequence.

Benefits
  • Fast, simple optimization.
  • Extremely high sensitivity and specificity using the PCR Enhancer Technology.
  • MasterAmp PCR PreMixes contain all components needed for successful PCR.
  • Generates PCR products suitable for both TA cloning and blunt-end cloning.

pcr_opt_1.jpg
 STEP 1. Find the optimal MasterAmp PCR PreMix. Do PCR with your template, primers and PCR enzyme in the 12 PreMixes.

A. A 75%-GC-rich ApoE gene region
 was amplified in total human genomic DNA using PreMixes A-L. MasterAmp PCR PreMix K produced optimal results in Step 1.
pcr_opt_2.jpg
 STEP 2. Use the same MasterAmp PCR PreMix to obtain reliable, consistent PCR of the same sequence.

B. Subsequent PCR
 of the ApoE gene with the same primer pair and MasterAmp™ PreMix K gave consistent results. Subsequent PCR results are consistent and reliable using the optimal PreMix in Step 1.

MasterAmp™ AmpliTherm DNA Polymerase; MasterAmp PCR Optimization Kits; MasterAmp Taq DNA Polymerase; MasterAmp Tfl DNA Polymerase. Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser's own internal research. No other patent rights (such as 5´ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Epicentre has two MasterAmp PCR Optimization Kits. Select the right kit for your DNA polymerase.
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