VeriFi™ Hot Start Polymerase - Long range, high fidelity PCR with AptaLock (Incl. dNTPs)

Art No | PB10.45

VeriFi™ Hot Start Polymerase - Long range, high fidelity PCR with AptaLock (Incl. dNTPs)

Choose size / variation:

from 130.00 CHF

PCRBIO HS VeriFi™ Polymerase is a versatile and robust proofreading enzyme with AptaLock™ hot start technology for highly precise PCR.

Description

Improved DNA binding and increased processivity give significant improvements in speed, yield and sensitivity while also increasing PCR success rates of long and challenging templates.

AptaLock™ hot start technology for maximised sensitivity and specificity
Greater success with long and/or GC or AT-rich templates (17.5kb and over)
High temperature cycling – up to 100°C denaturation to better separate GC-rich sequences
100x higher fidelity than Taq DNA polymerase
Room temperature setup
Reaction mix stability for up to 24 hours both before and after PCR run
Generates blunt-end PCR products
Also available as a 2x ready mix with the option of a red dye for direct gel loading

VeriFi polymerase came to my rescue when I struggled with amplifying large vectors using regular xxx polymerase.

Its high fidelity and good yield made a noticeable difference. Further, the same polymerase can be used for amplifying fragments from genomic DNA, which is known to inhibit a lot of polymerases, therefore making it a true all-in-one solution. With over half a decade of experience, I confidently recommend it to anyone tackling large vectors.

Trust me, VeriFi will make your lab life a whole lot easier!

Gopinath Chattopadhyay, Ph.D.

MBU, Indian Institute of Science, India

Current Position: Postdoc, IEU, University of Zurich, Switzerland

PCRBIO HS VeriFi™ Polymerase is a single enzyme derived from Pfu DNA polymerase for its 3’-5’ exonuclease (proofreading) activity. Proprietary mutations improve DNA binding and increase processivity when compared to its native form, resulting in shorter extension times, higher yields and the ability to amplify longer and more difficult targets. PCRBIO HS VeriFi™ Polymerase is able to amplify eukaryotic genomic templates in excess of 17.5kb, and longer for simpler DNA templates. Extension times of 30s per kb are recommended for most applications.

PCRBIO’s innovative AptaLock™ technology uses a proprietary aptamer-like molecule that reversibly inhibits both the 3’-5’ exonuclease activity and 5’-3’ polymerase activity of the enzyme below 40°C. This unique hot start molecule prevents primer dimer formation and non-specific amplification to maximise the sensitivity and specificity of your PCR. This feature makes PCRBIO HS VeriFi™ Polymerase highly suitable for multiplexing and enables reactions to be set up at room temperature. The reaction mix is stable both before and after the PCR run for up to 24 hours, meaning greater ease of use for high throughput assays.

The enhanced accuracy of PCRBIO HS VeriFi™ Polymerase gives extremely low error rates and fidelity that is approximately 100 times higher than Taq DNA polymerase. The enzyme is ideal for applications where superior accuracy is required, such as cloning, site-directed mutagenesis and sequencing.

The enzyme is provided with an advanced buffer system including dNTPs, Mg and enhancers, enabling high fidelity PCR of a wide range of targets and fragment sizes regardless of GC or AT content. For the most difficult templates, such as highly GC-rich sequences or those with complex secondary structures, users have the option of adding the VeriMax Enhancer to the reaction mix for improved PCR performance.

PCRBIO HS VeriFi™ Polymerase is also available as a convenient 2x ready mix containing all reaction components apart from primers and template. Our 2x red mix contains a red dye suitable for direct loading and tracking during agarose gel electrophoresis.

Applications

High fidelity PCR
Long PCR
Multiplex and high throughput PCR
Site-directed mutagenesis
Cloning
Sequencing

Storage

On arrival, products should be stored between -30 and -15°C. If stored correctly the kit will retain full activity for 12 months.

Material Safety Data Sheet (PDF)

Product manual (PDF)

FAQs

Figure 1. Superior performance in multiplex reactions

10-plex PCR using lambda phage genome (6 targets) and mouse genome (4 targets) at different annealing temperatures (A: 63.0°C, B: 61.5°C, C: 60.5°C). The starting template amount is 1pg lambda DNA and 1ng mouse gDNA. Amplicon lengths are between 139bp and 962bp. Reactions were set up using master mix formats following manufacturers’ recommendations. Cycling conditions were 95°C 2 min, 40 cycles of 95°C 15 sec, annealing A to C 30 sec, 72°C 90 sec. L: PCRBIO Ladder III. P: reference pool of single products.

PCRBIO HS VeriFi™ Mix displays greater sensitivity and specificity in multiplex when compared to leading competitors.

Figure 2. Successful PCR across a broad range of GC and AT content

Amplification of 13 targets with GC content ranging from 28.7% to 83% using PCRBIO HS VeriFi™ Mix. The starting template amount is 30ng mouse cDNA. Band size is between 99bp and 274bp. Cycling conditions were 98°C 5 min, 40 cycles of 98°C 15 sec, annealing between 54°C and 62°C (depending on target) 15 sec, 72°C 30 sec. L: PCRBIO Ladder III.

PCRBIO HS VeriFi™ Mix is able to amplify templates across a broad range of GC and AT content.

Figure 3. Increased PCR success rates and consistency with complex targets

Amplification of a 13.5kb fragment of the human ß-globin gene at different annealing temperatures (A: 68.5°C, B: 66.0°C, C: 63.0°C, D: 60.5°C). The starting template amount is 30ng human genomic DNA. GC content is 37%. Reactions were set up using master mix formats (apart from Takara’s PrimeSTAR® GXL DNA Polymerase) and following manufacturers’ recommendations. Cycling conditions were 95°C 2 min, then 30 cycles of 95°C 15 sec, annealing 15 sec, 72°C 12 min.

PCRBIO HS VeriFi™ Mix displays higher yield and specificity compared to leading competitors. The PCRBIO mix also shows greater consistency and versatility across the annealing temperature range.

Increased PCR success rates with high CC templates:

Amplification of 600bp fragment of ATXN2 gene with CC content of 71%. The starting template amount was 5ng mouse genomics DNA. Reactions were set up using master mix formats (apart from Takara PrimeSTAR GXL Polymerase) and following manufactures’ recommendations. Cycling conditions were 99°C 5 min., then 30 cycles off 99°C 30 sec., 60°C 30 sec., 72°C 45 sec.

L: PCRBIO Ladder III.

PCRBIO HS VeriFi™ Mix amplifies this hight CC sequences with greater sensitivity and specificity compared to leading competitors.

PCRBIO’s innovative AptaLock™ technology

uses a proprietary aptamer-like molecule that reversibly inhibits both the 3’-5’ exonuclease activity and 5’-3’ polymerase activity of the enzyme below 40°C. This unique hot start molecule prevents primer dimer formation and non-specific amplification to maximise the sensitivity and specificity of your PCR.

Available sizes:

PB10.45-01	1 x 50μL \| 100 Units
PB10.45-05	1 x 250μL \| 500 Units

Supplier

PCR Biosystems
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Specifications

PCRBIO HS VeriFi™ Polymerase (2u/μL)	1 x 50μL \| 100 Units 1 x 250μL \| 500 Units
5x PCRBIO VeriFi™ Buffer	1 x 1.7mL \| 100 Units 3 x 1.7mL \| 500 Units
10x VeriMax Enhancer	1 x 1.7mL \| 100 Units 2 x 1.7mL \| 500 Units
Reaction Volume	50μL