VeriFi® Library Amplification Mix for NGS

Features:

• Low GC bias, ideal for high GC/AT targets
• More unique reads per NGS dataset for superior data quality
• AptaLock™ hot start technology for maximum sensitivity and specificity
• 100x higher fidelity than Taq DNA polymerase
• Room temperature setup
• 2x ready mix for minimal pipetting
• Resistant to blood and chemical inhibitors

Application

• NGS library amplification
• Whole genome sequencing
• RNA-Seq
• Multiplex and high throughput PCR
• GC/AT rich target sequencing
• Metagenomic studies

VeriFi® Library Amplification Mix

VeriFi® Library Amplification Mix was created as a tool for researchers conducting next generation sequencing (NGS) studies. A key part of PCR-based NGS workflows is the amplification step during which a DNA or cDNA (in the case of RNA-Seq studies) library has been modified to contain appropriate adaptors for the intended sequencing platform and needs to be amplified before proceeding with the sequencing step.

To ensure accuracy of the final sequencing data, proofreading polymerases used in the amplification step of an NGS pipeline must have certain key characteristics, which are:

• High Fidelity, to minimise PCR errors in nucleotide incorporation.
• Strong multiplexing capability, to amplify varying fragment lengths in a library.
• Low sequence-dependent bias, particularly when dealing with complex (GC/AT-rich, or repetitive) sequences.

Additional features can also help improve library amplification mix performance, depending on the type of NGS study being undertaken.  VeriFi® Library Amplification Mix is engineered with these features in mind. It is powered by our market leading VeriFi® Polymerase, with AptaLock™ hot start technology, in a novel 2x mix format. The mix contains reaction buffer, dNTPs, Mg²⁺, and enhancers that minimise GC-bias and enable reliable library amplification for Illumina sequencing platforms. Amplified libraries can easily be quantified using NGSBIO Library Quant Kit Blue for Illumina prior to sequencing.

Better quality NGS data

VeriFi® Library Amplification Mix was externally validated using Illumina-based sequencing in a blind comparison with three market-leading competitors. VeriFi® Library Amplification Mix consistently gave 2% more uniquely mapped reads than competitors for three different microbial genomes. Depending on sequencing platform being used, this small percentage can translate to 80,000 to 100,000,000 more unique reads. This leads to greater target sequence coverage and more accurate quantitative information, making sequencing experiments more cost-effective and resulting data more informative.

Reduced GC bias during NGS library amplification

Novel buffer composition makes VeriFi® Library Amplification Mix ideal for complex templates. Our research team has challenged this enzyme in PCRs on numerous difficult templates, demonstrating minimal bias during amplification at extremes of GC content ranging from 30% – 70%. With consistently better performance than all major competitors, these results ensure cutting edge performance of the mix regardless of the target sequence, to give you confidence when undertaking costly and laborious NGS experiments. In combination with a higher number of unique reads, discussed above, this feature makes VeriFi® Library Amplification Mix perfect for conducting metagenomic analyses.

AptaLock™ hot start technology

VeriFi® Library Amplification Mix is enhanced by PCRBIO’s innovative AptaLock™ hot start technology, a proprietary aptamer-like molecule that reversibly inhibits both the 3’-5’ exonuclease activity and 5’-3’ polymerase activity of the enzyme at ambient temperatures. The unique hot start molecule prevents primer dimer formation and non-specific amplification to maximise PCR sensitivity and specificity. This ensures amplification reactions can be set up at room temperature.