Enabling sequencing of many DNA strands at the same time, Next-Generation Sequencing (NGS), or initially called ''massively parallel sequencing'', has revolutionized genomic research and played a significant role when applied in academic research, clinical diagnostics, agricultural studies, forensic science and other fields.
As a leading NGS products provider, iGeneTech offers full spectrum of NGS solution to meet your needs. To decode the mystery of life, we design and manufacture panels of your specific interests. To streamline the process of library preparation, we provide cost-effective reagent kits together with simple and optimized workflow. To achieve massive production, we have the automation solution applied to various experiment steps. From each individual module to the entire portfolio, we are here to offer high quality products to explore the unknown.
Target Enrichment approaches are commonly used in next-generation sequencing (NGS) workflows to effectively save on sequencing costs by eliminating genes or regions that are not of interests. Through target enrichment sequencing, one can focus on the specific regions and obtain greater depth enough for subsequent analysis with less time and expense when compared to whole genome sequencing.
The two most popular techniques for NGS target enrichment are hybridization capture-based and multiplex PCR-based enrichment (amplicon sequencing), both of which are essential and have been widely used in many fast-growing clinical, academic and industrial applications. To meet the increasing demand for different targets of interests, iGeneTech has developed varies of proven panels based on TargetSeq® Hybridization Capture Sequencing Technology, BisCap® Methyl Hybridization Capture Sequencing Technology as well as MultipSeq® Multiplex Amplicon Sequencing Technology, where customization is also available to build your own panels.
iGeneTech hybridization capture-based and multiplex PCR-based enrichment can also be applied to Sanger sequencing and third-generation sequencing. Please Contact Us for any questions and inquires.
For qualified samples, most of the human DNA TargetSeq® panels can be evaluated by using the criteria of Coverage rate (%) > 99%, Target reads capture rate (%) > 50%, T 20% X coverage rate (%) > 97%. For TargetSeq® panels of other species, the performance might vary due to the differences of reference genome, target regions and strains.
We provide different design strategy to detect CNV according to its different sizes. For CNVs over 10 kb, a strategy called CNV backbone can be used by including high MAF SNPs at the intervals based on the target CNV sizes. For exon level CNVs (100 to 1000 bp), panels can be designed by covering all the related exon regions. The AIExome® Human Exome Panel V3 – Inherit includes the specially designed CNV backbone, which can be used for CNVs over 500 kb at whole genome level and CNVs over 50 kb at curated regions.
There are some regions in the genome that we classify as hard-to-capture regions, including high GC content sequences, repetitive sequences and homologous sequences. Based on the evaluation result, some of these target regions might be dropped as they might affect the overall performance or introduce errors. The probe coverage of most designs will be over 99% and the missed regions will be specified in probeMissed.bed.
No. DNA panels are designed based on the reference genome, while the RNA panels are designed based on the transcripts. RNA panels normally include additional probes at the edge of exons in order to increase the detection rate of fusions.
In our design report, we provide different versions of bed files for different analysis. Loci.bed and ProbeCov.bed are the bed files for target regions and the regions covered by actual probes, respectively. Please select the appropriate bed file according to your requirements. Please be noted that if the target is a point mutation or the panels includes the genome fixed/novel patches, the performance evaluated by using Loci.bed and ProbeCov.bed will be significantly different.
For human panels, TargetSeq® panel uses the 3X tiling design strategy based on thermodynamic stability, and a specifically optimized multi-factor strategy for hard-to-capture regions and different variation types to improve the overall performance and the detection rate of key markers. For other species, based on the self-developed algorithm, we will perform the evaluation of the genome first and design the panels using different tiling strategy to ensure the balance of the performance and the cost.
CN