|VeraSeq DNA Polymerase 2.0
VeraSeq high-fidelity DNA polymerase 2.0 is an engineered, ultra-thermostable polymerase designed to maximize the speed, accuracy, and length of DNA synthesis during sequencing template preparation. The result is a novel enzyme that can extend a kilobase of sequence in 15 seconds and with accuracy 50 times higher than Taq DNA Polymerase.
Source of Protein
A recombinant E. coli strain carrying the engineered VeraSeq gene.
20 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
pH 7.4 @ 25°C
5X VS Buffer 2 (B7102)
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.
Double-Stranded Endonuclease Activity
A 50 µL reaction containing 0.5 µg of pBR322 DNA and 10 µL of enzyme solution incubated for 4 hours at 37°C resulted in no visually discernible conversion to nicked circular DNA as determined by agarose gel electrophoresis.
E.coli 16S rDNA Contamination Test
Replicate 5 µL samples of enzyme solution were 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. The acceptance criterion for the test is the threshold cycle count (Ct) produced by the average of 3 replicate no template control samples. Based on the correlation between the no template control Ct values, and standard curve data, the detection limit of this assay is <10 copies genome/sample.
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.
||-25 to -15°C
||120 U = no conversion
|E.coli DNA Contamination
||150 U < 10 copies
* For a detailed summary of assay conditions and data,
refer to the Quality Controls Analysis section below
Quality Control Analysis:
Unit Characterization Assay
Specific activity was measured using a 2-fold serial dilution method. Dilutions of enzyme were made in 1X reaction buffer and added to 50 µL reactions containing activated calf thymus DNA; 25mM TAPS (tris-[hydroxymethyl]-methyl- amino-propanesulfonic acid, sodium salt),pH 9.3 (at 25°C); 50mM KCl; 2mM MgCl2; 1mM ß-mercaptoethanol; 200µM each dATP, dGTP, dTTP; and 100µM *Α-32P]-dCTP (0.05 to 0.1 Ci/mmole). Reaction vessels were mixed and incubated at 74°C for 10 minutes.
Protein Concentration (OD280) Measurement
A 2.0 µL sample of enzyme was analyzed at OD280 using a Nanodrop ND-1000 spectrophotometer standardized using a 2.0 mg/ml BSA sample (Pierce Cat #23209) and blanked with product storage solution. The observed average measurement of 3 replicate samples was converted to mg/mL using an extinction coefficient of 134,130 and molecular weight of 97,697 Daltons.
SDS-Page (Physical Purity Assessment)
2.0 µL of concentrated enzyme solution was loaded on a denaturing 4-20% Tris-Glycine SDS-PAGE gel flanked by a broad-range MW marker and 2.0 µL of a 1:100 dilution of the sample. Following electrophoresis, the gel was stained and the samples compared to determine physical purity. The acceptance criteria for this test requires that the aggregate mass of contaminant bands in the concentrated sample do not exceed the mass of the protein of interest band in the dilute sample, confirming greater than 99% purity of the concentrated sample.