Human Long-Amplicon Sequencing

Amplicon sequencing is a highly targeted approach in which PCR products (also known as amplicons) are sequenced ultra-deep to efficiently identify and characterize variants, allowing researchers to study genetic variation in specific genomic regions. However, the correct phasing of genetic variations using amplicon sequencing by short-read sequencing technologies remains challenging. Long-read next-generation amplicon sequencing show promise as a powerful tool for studying complete genes or genomes of complex and diverse populations. Based on Single Molecule, Real-Time (SMRT) sequencing and its advanced Sequel sequencing system, and years of experience in the study of human genetic variation, CD Genomics is proud to offer professional, reliable, and cost-effective human long-amplicon sequencing services to meet the project needs of our clients worldwide.

Using PacBio circular-consensus-sequencing (CCS) analysis, we can generate high accuracy single-molecule consensus sequences of large genomic regions (amplicons ranging in size from several hundred base pairs to 20 kb with a low error rate and chimera rate). Amplicon and genomic sequences up to 12 kb in length can yield the highest quality single-molecule sequence data. Our services can provide high fidelity (HiFi) individual reads containing phased variants and breakpoints and have many applications, such as the detection of disease-causing mutations and the investigation of genetic disorders.

The Pipeline of Long Amplicon Analysis (LAA) using SMRT Sequencing

Sample Requirements

1. High-quality nucleic acids: 200ng or more.
2. Since we offer amplicon preparation services, we also accept other samples, such as DNA fragments, plasmids, cell pellets, intestinal contents, saliva, skin, and other samples.

Data Analysis

-Circular consensus sequence (CCS) analysis.
-Long amplicon analysis (LAA).
-Characterization of genetic complexity, including structural variation (SV), rare single nucleotide polymorphisms (SNPs), insertion-deletion mutations (InDel), copy number variation(CNV), microsatellites, and haplotypes.
-Minor variant analysis.

Some Application of Our Human Long-Amplicon Sequencing 

• Immune repertoire sequencing, such as antibody screening sequencing
• Determination and analysis of important pharmacogenetic genes.
• Discovery, validation, and screening of somatic/complex variants.

The diversity and importance of phasing for some loci in the human genome make it difficult to analyze them with traditional sequencing methods. Since PacBio SMRT sequencing can provide long reads with high consensus accuracy (>99% single-molecule read accuracy), combined with long amplicon sequencing, it is suitable for analyzing difficult genomic loci such as the genes from the Major Histocompatibility Complex (MHC). In addition, long amplicon analysis has proven to have good applications in the study of human leukocyte antigen (HLA) amplicons. With the higher output of the PacBio Sequel system, the cost per sample will be further reduced, making long amplicon sequencing using SMRT sequencing a suitable alternative to targeted short-read length methods. CD Genomics is a leading global life sciences company, and we continue to expand our services and improve our existing resources to help scientists accelerate their research and support their careers. One of our goals is to generate reference-quality assemblies in diverse populations to better help scientists understand the complexity of human health and disease. For more information on how we can help you, please feel free to contact us.