With the introduction of the Pwo Master, Roche Applied Science now offers a new, ready-to-use proofreading master mix for the robust amplification of fragments up to 3 kb, providing the highest possible fidelity during PCR synthesis.

Pwo Master is a premixed double-concentrated formulation that contains PwoSUPER YIELD DNA Polymerase, a new optimized buffer system, and PCR-Grade Nucleotides. After simply adding the template DNA and the primers and adjusting the final volume to 50 l with PCR-Grade Water (provided with the Pwo Master), the reaction can be started.

The reaction setup of a large number of samples for PCR analysis requires a multitude of pipetting and handling steps. Every step in PCR is critical to the quality of the final result; pipetting and handling errors, as well as assay contamination, can negatively impact the experimental outcome, leading to false-negative or false-positive results. As a result, PCRs must often be repeated, resulting in wasted effort and reagent expense. The availability of master mixes simplifies the PCR setup and circumvents many problems.

Enzymology of Pwo Master

The core component of the Pwo Master is the proofreading polymerase Pwo DNA Polymerase. Pwo DNA Polymerase is a highly processive 5′3′ DNA polymerase that possesses 3′5′ exonuclease activity, also known as proofreading activity. This proofreading activity results in an approximate 18-fold increase in the fidelity of DNA synthesis compared with Taq DNA polymerase.

PwoSUPER YIELD DNA Polymerase combines Pwo DNA Polymerase with a new optimized buffer system which enhances the enzymatic properties of the enzyme, resulting in higher yields and additional robustness of the amplification reaction without changing the fidelity of DNA synthesis.

Advantages of Pwo Master



Avoid the need to determine the optimal buffer conditions for your proofreading PCRs

• The Mg2+ concentration of the master mix in combination with the specially formulated buffer system allows the amplification of most fragments up to 3 kb with high yield and specificity (Figure 1), and the high fidelity you expect from a proofreading polymerase (Figure 2).
• There is no need to titrate the optimal ionic conditions for a given assay or to perform cumbersome adjustments of the reagent compositions (Figure 1).
  
  

Improve the reproducibility and consistency of your proofreading PCRs

• The number of pipetting steps during PCR setup is reduced.
• The risk of contaminating the reaction mix during PCR setup is reduced.
• The analysis of your results is straightforward.
  
  

Save time the Pwo Master is ready when you are

• The Pwo Master is stored at 2 8C and can be used immediately; no freezing or thawing is necessary.
  
  

Contents of Pwo Master

• Pwo Master: 10 vials of double-concentrated master mix, 10 reactions per vial. Each vial contains 25 units PwoSUPER YIELD DNA Polymerase, reaction buffer with 4mM MgCl2 and PCR-Grade Nucleotides (dATP, dCTP, dGTP, dTTP, each 0.4 mM) in a total volume of 250 l.
• PCR-Grade Water: four vials of 1 ml each.
  
  

β-Glucuronidase from E. coli K12 New improved quality!

Roche Applied Science recently introduced an improved β-Glucuronidase (GRD) formulation which replaces the previous product. The new quality exhibits higher purity and specific activity compared with products previously on the market (Figure).



GRD hydrolyses β-glucuronic acid (β-GlcA) esters (β-glucuronosides) of a variety of compounds, including steroids, aryl alcohols, drugs, and metabolites. GRD also hydrolyses polysaccharides that contain β-GlcA linked to other sugars. GRD is highly specific for the carbohydrate portion of the molecule but has very little specificity for the molecule conjugated to β-GlcA. The main research applications are:

• Analysis of steroid conjugates
• Doping analysis (horses)
• Drug and alcohol testing
• Tests in pathology labs
  
  

The performance in steroid analysis and doping analysis was demonstrated by Ute Mareck, German Sport University, Cologne, Germany, and Thierry Rousseau, National Doping Testing Laboratory (LNDD), Chtenay- Malabry, France.




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Related biology technology :

1. Efficient and Reliable PCR Setup Using Eppendorf MasterMix
2. Comparing Fidelity and Performance of Proofreading PCR Enzymes