What are the main applications for the Equinox Library Amplification Kit?
Our Equinox Library Amplification Kit has been specifically optimized for efficient NGS library amplification from a wide range of template amounts (0.1 pg – 500 ng) up to 1 kb in length. These NGS applications include:
- Low-frequency variant detection NGS assays, including those utilizing challenging samples such as FFPE and cfDNA
- Hybridization capture workflows
- Single-cell analysis
- Whole genome sequencing (WGS)
- Amplicon sequencing
- ChIP-Seq, ATAC-Seq, and associated epigenetic applications
- Illumina and non-Illumina sample preparation workflows
What type of enzyme is in the Equinox Library Amplification Kit?
Equinox DNA Polymerase is a proprietary B family proofreading polymerase variant developed specifically for NGS applications to deliver excellent fidelity, uniform sequence coverage, and high library complexity.
Can the Equinox Library Amplification Kit amplify longer templates?
What is the hot start functionality of Equinox and how well does it work?
Does Equinox tolerate uracil residues?
What are the cycling recommendations for the Equinox Library Amplification Kit?
Table 1: Thermocycler program settings
|Step||Temperature (°C)||Time (sec)||Cycles|
|Denaturation||98||15||See Table 2|
1 An annealing temperature of 60°C is recommended for standard Illumina® “P5” and “P7” primers (P5:
CAAGCAGAAGACGGCATACGAGAT). For other amplification primers, the optimal annealing temperature should be determined empirically in an annealing temperature gradient (55°C – 70°C) experiment.
2 Longer extension times may be employed to ensure efficient amplification of longer-insert libraries. A 30 sec extension is sufficient for libraries with a mode fragment size up to 500 bp; a 45 sec extension time is recommended for libraries with mode fragment sizes >500 bp. The optimal condition for each application may need to be determined empirically.
Table 2: Recommended PCR cycle numbers based on DNA input
|Adapter-ligated DNA (ng)||PCR cycles to generate|
|40 nM library||1 µg library|