Qiagen, P7610L, VeraSeq PCR Mix (2x, 250 reactions)

250 reactions (evaluation pack) of 2x VeraSeq PCR Mix (1 x 6.25 mL)

Features

- 2x master mix based on VeraSeq™ 2.0 (cat. no. P7511L)
- 50x greater fidelity than Taq DNA Polymerase
- Ultra-thermostable polymerase
- Extends 1 kb in 15 seconds
- Strong proofreading activity (3'→5' exonuclease activity)

Product Details

VeraSeq™ PCR Mix is a premixed, ready-to-use 2x solution containing VeraSeq™ 2.0 High-Fidelity DNA Polymerase (P7511L) , dNTPS, MgCl 2 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

Test: Specification
Functional Assay: Amplification of 500 bp fragment from genomic DNA

- 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

Polymerase properties

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

Component: Volume (µL)
Sterile H 2 O: 20, variable
2x VeraSeq PCR Mix: 25
PCR Primer Cocktail: 5
Library DNA*: Variable

Step: Temperature
Initial denaturation: 98°C
Denaturation Annealing Extension: 98°C 60°C 72°C
Final extension: 72°C 4°C

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 50 µL)

* Total reaction volumes of library DNA and water should be adjusted to achieve a final reaction lolume of 50 µL. 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.

* 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.

Contamination Tests

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.

Applications

- High-fidelity DNA amplification
- Cloning
- Synthetic biology

This product is available for molecular biology applications such as: