Cat. No. / ID: P7610L
VeraSeq™ PCR Mix is a premixed, ready-to-use 2x solution containing VeraSeq™ 2.0 High-Fidelity DNA Polymerase (P7511L), 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.
Polymerase properties
Test | Specification |
Functional Assay | Amplification of 500 bp fragment from genomic DNA |
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.
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)
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 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.
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.
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.
This product is available for molecular biology applications such as: