For Illumina® Platforms
Targeted sequencing, which allows you to focus on sequences and region of interest, encapsulates two different techniques: a broad, target capture via a hybridization method or a more focused, PCR-based amplicon panel design. Among its most notable benefits compared to other methods in NGS such as WGS or WES, is the fact that because less genomic sequence is being interrogated, only target regions of interest are sequenced. This means that by focusing the sequencing real estate of the flow cell to only regions of interest, genomic sequences can be enriched anywhere from 100 to a 1 million times compared to DNA-sequencing. This is critical for some applications where the typical 10x or 100x coverage obtained by DNA-sequencing or even large exome-sequencing may not be sufficient, such as for the detection of low level somatic variants that may require 3,000x coverage or more to be detected at a frequency of 5% or less. Similarly, great confidence can be got with deep sequencing via NGS than Sanger sequencing for these low-frequency variant calls.
PerkinElmer offers a complete line of NEXTFLEX® library preparation kits to meet your targeted sequencing needs.
Centered around the NEXTFLEX® Human Whole Exome Sequencing Panel and NEXTFLEX® Core Exome Sequencing v2 panel, PerkinElmer’s standardized and tested targeted capture workflows ensure time and cost savings to labs that are looking to establish genetic analysis or expand their current test menu.
Amplicon panels, which excel over target-capture hybridization methods that cast a broader net into the genome, are the technology of choice for small target panels ranging anywhere from 5 – 50 kb of sequence. They are also much cleaner, more rapid, and simpler to carry out on the bench when compared to any method requiring hybridization due to the fact that it is PCR-based. They also tend to be the most cost-efficient for sequencing as it requires the least amount of sequencing per sample for most applications, such as in the detection of germline mutations that only typically require coverage of approximately 50x – 500x. Similarly, as a PCR-based technology, different tactics can be employed that would not be viable using other sequencing methods to tackle the resolution of difficult genomic regions, such as those with high AT-rich or GC-rich sequences and paralogues/pseudogenes. Amplicon panels can sometimes be used in the context of targeted re-sequencing due to their ability to interrogate regions that may have been missed or insufficiently covered by WGS or WES. Their utility as an alternative option to reflex Sanger sequencing of problematic regions has also been adopted by some in the field as well.
16S & 18S Sequencing Kits
16S and 18S rRNA sequencing are useful and cost-effective NGS tools for microbial sequencing for metagenomic studies comparing different species of bacteria or for eukaryotic cells (18S ITS) present in a given sample without the need for cultures. 16s rRNA is a widely used gene marker for genus and species identification in bacteria and archaea for its minuscule rate of substitution in the highly conserved sequences outside of the hypervariable regions. 18S rRNA is routinely investigated in the phylogenetic study of fungi as it contains more hypervariable domains than 16S. The speed and economy associated with amplicon sequencing is maximized with this approach catered to population genetics and microbial profiling for interrogating the phylogeny and taxonomy of samples from complex microbiomes.
NEXTFLEX® Variant-Seq™ SARS-CoV-2 Kit
The NEXTFLEX® Variant-Seq™ SARS-CoV-2 kit has optimized the identification and tracking of SARS-CoV-2 variants by quickly, easily, and affordably identifying all mutations in a SARS-CoV-2 sample using amplicon sequencing. Sample preparation has been simplified, throughput has been increased, and multiple automation solutions offer flexibility based on your lab’s needs.