VeraSeq PCR Mix

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VeraSeq PCR Mix (2x, 250 reactions)

Cat. No. / ID: P7610L

For 250 reactions (evaulation pack) of premixed, ready-to-use 2x solution containing VeraSeq 2.0 High-Fidelity DNA Polymerase, dNTPs, MgCl₂ and reaction buffer.

The VeraSeq PCR Mix (2x, 250 reactions) is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

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Certain applications in which this product can be used may be covered by patents issued and applicable in the United States and abroad. Purchase of this product does not include a license to perform any patented application; therefore it is the sole responsibility of the users of the product to determine whether they may be required to engage a license agreement depending upon the particular application in which the product is used. This product is licensed from Bio-Rad Laboratories, Inc., under U.S. Pat. Nos. 6,627,424,7,541,170,7,560,260 and corresponding patents in other countries for use solely in DNA sequencing, DNA micro-array, and conventional PCR applications, including pre-amplification steps that are required for such applications, in the life science research and in-vitro diagnostics fields but not real-time PCR or digital PCR.

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Features

2x master mix based on VeraSeq 2.0 (P7511L)
Ultra-thermostable polymerase
Extends 1 kb in 15 seconds
50x greater fidelity than Taq DNA Polymerase
Strong proofreading (3'-5' exonuclease)

Product Details

VeraSeq PCR Mix is a premixed, ready-to-use 2x solution containing VeraSeq 2.0 High-Fidelity DNA Polymerase, dNTPS, MgCl₂ and reaction buffer at optimal concentrations to maximize the speed, accuracy and length of DNA synthesis. The formulation provides efficient, high-fidelity DNA amplification, cloning, and synthetic biology applications.

Performance

Polymerase properties

Storage temperature: –25°C to –15°C
Extension rate: 15 second per kb at 72˚C 
Proofreading (3'-5' exo): Yes, strong 
Nick-translation (5'-3' exo): No 
Fidelity: >50x higher than Taq DNA Polymerase 
Strand displacement:  No 
Thermostability: Highly thermostable 
Able to extend an RNA primer: No
Extends from a nick: No 
Generate blunt end products: Yes 
Uracil read through: No

Test	Specification
Functional Assay	Amplification of 500 bp fragment from Genomic DNA

Principle

Source of recombinant enzyme protein
The protein is produced by a recombinant E. coli strain carrying the engineered VeraSeq 2.0 gene.

Unit definition
One unit is defined as the amount of enzyme required to incorporate 10 nmoles of dNTPs into acid-insoluble form at 74°C in 30 minutes.

Procedure

Protocol
General precautions should be taken when setting up a PCR, including setting up the reaction on ice, adding master mix last, gently pipetting, thorough mixing and a quick centrifugation. The following procedure can be used as a guideline. Reactions may need to be optimized individually.

Reaction setup (for 50µl)

Component	Volume (µl)	Final concentration
Sterile H₂O	20, variable	n/a
2x VeraSeq PCR Mix	25	1x
PCR Primer Cocktail	5	0.5 µM each
Library DNA*	Variable	n/a

* Total reaction volumes of library DNA and water should be adjusted to achieve a final reaction lolume of 50µl. If the reaction volume needs to be >50 µl, the volume of the 2x Master Mix should be adjusted so that it constitutes 50% of the final reaction volume.

Typical cycling conditions*

Step	Temperature	Time	Cycles
Initial denaturation	98°C	30 seconds	1
Denaturation Annealing Extension	98°C 60°C 72°C	10 seconds 30 seconds 30 seconds	To be determined by user^†
Final extension	72°C 4°C	300 seconds hold	1

* Cycling conditions may need to be optimized, depending on the amplicon of interest.
^† Number of cycles is dependent on the amount of input DNA and other specific sequence analysis requirements.

Quality control analysis
Functionality of 2x VeraSeq PCR Mix is assessed by its ability to amplify a 500 bp fragment from genomic DNA. Following PCR the 500 bp fragment was visualized by agarose-gel electrophoresis.

VeraSeq PCR Mix was tested prior to assembly and found to be free of contaminating endonucleases. Enzyme purity was >99% as determined by SDS-PAGE and negligible E. coli genomic DNA contamination was confirmed by qPCR. Specific activity was verified pre and post dilution.