Cat. No. / ID: P7511L
VeraSeq™ 2.0 High-Fidelity DNA Polymerase is an engineered, ultra-thermostable polymerase that delivers industry-leading
speed, fidelity and robustness to PCR amplification. The novel enzyme can extend a kilobase of sequence in 15 seconds and with
an accuracy 50 times higher than Taq DNA Polymerase.
Supplied in:
20 mM Tris·HCl, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, Stabilizer and 50% glycerol; pH 7.4 at 25°C.
Supplied with:
5x VeraSeq™ Buffer II (B7102L) and 5x VeraSeq™ GC Buffer (B7130L).
You can also ask us about low glycerol or hot-start formulations.
Polymerase properties
Extension rate: 15 seconds 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 | Units Tested | Specification |
Purity | n/a | >95% |
Specific activity | n/a | 100,000 U/mg |
Double-stranded endonuclease | 120 U | No conversion |
E. coli DNA contamination | 150 U | <10 copies |
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.
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 polymerase last, gentle 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 | Variable | |
5x VeraSeq™ Buffer II or 5x VeraSeq GC Buffer | 10 | 1x |
10 mM dNTP mix | 1 | 200 µM each |
Primer 1 | Variable | 0.2 µM |
Primer 2 | Variable | 0.2 µM |
DNA template | Variable | See note 4 |
VeraSeq™ 2.0 DNA Polymerase | 0.5 | 1 U |
Total reaction volume can be adjusted as needed.
Typical cycling conditions
Step | Temperature | Time | Cycles |
Initial denaturation | 98°C | 30 seconds | 1 |
Denaturation Annealing Extension |
98°C Varies 72°C |
5–10 seconds 10–30 seconds 15–30 seconds/kb |
15–35 |
Final extension | 72°C 4°C |
5–10 mininutes Hold |
1 |
Cycling conditions may need to be optimized, depending on the amplicon of interest |
Usage Notes:
Complexity | Source example | Guideline |
---|---|---|
Low | Plasmid, virus, BAC | 1 pg – 10 ng |
High | Genomic DNA | 50–250 ng |
Quality control analysis
Unit activity was measured using a twofold serial dilution method. Dilutions of enzyme were made in 1x reaction buffer and added
to 50 µl reactions containing activated calf thymus DNA; 25 mM TAPS (tris-[hydroxymethyl]-methyl-amino-propanesulfonic acid, sodium salt), pH 9.3 at 25°C; 50 mM KCl; 2 mM MgCl2; 1 mM β-mercaptoethanol; 200 µM each dATP, dGTP, dTTP; and 100
µM [3H]-dCTP (0.075 Ci/mmole). Reaction vessels were mixed and incubated at 74°C for 10 minutes.
Protein concentration is determined by OD280 absorbance.
Physical purity is evaluated by SDS-PAGE of concentrated and diluted enzyme solutions followed by silver-stain detection. Purity is
assessed by comparing the aggregate mass of contaminant bands in the concentrated sample to the mass of the band corresponding
to the protein of interest in the diluted sample.
Double-stranded endonuclease activity is determined in a 50 µl reaction containing 0.5 µg of plasmid DNA and 10 µl of enzyme
solution incubated for 4 hours at 37°C.
E. coli 16S rDNA contamination is evaluated using 5 µl replicate samples of enzyme solution that are denatured and screened in
a TaqMan qPCR assay for the presence of contaminating E. coli genomic DNA using oligonucleotide primers corresponding to the
16S rRNA locus.
This product is available for the following molecular biology applications: