Next Generation Sequencing

Creative Biogene's comprehensive NGS services for clients. We sequence millions of fragments in massively parallel fashion, increasing speed and accuracy while reducing sequencing costs. Additionally, we help our biopharma and biotech partners develop, validate and execute NGS assays to comply with GLP or cGMP regulations. We are committed to helping you unravel disease-causing factors from your genetic material through NGS, improving many aspects of scientific research and advancing the field of personalized medicine.

Fig.1 Basic scheme of a next generation sequencing experiment.

What Is Next Generation Sequencing?

Next-generation sequencing (NGS) is a massively parallel sequencing technology that provides ultra-high throughput, scalability, and speed. This technique is used to determine the sequence of nucleotides throughout the genome or in target regions of DNA or RNA. NGS technology has revolutionized genomics and molecular biology research by enabling faster and cheaper sequencing of DNA and RNA compared to previously used Sanger sequencing, enabling laboratories to perform a variety of applications at unprecedented levels and study biological systems. Today's complex genomics problems require a depth of information beyond the capabilities of traditional DNA sequencing technologies. NGS fills this gap and becomes an everyday tool to address these issues.

Our Next-generation Sequencing Workflow

• Sample Extraction

The NGS process begins by extracting nucleic acid (ie, DNA, total RNA, mRNA, or chromatin) that will be used for sequencing. Depending on the purpose of the experiment, genetic material can be extracted from a variety of biological samples, including blood, cultured cells, biopsies, tissue sections and urine, as well as microbial or plant specimens.

• Library Preparation

Prepare DNA or RNA samples for compatibility with sequencers, typically by fragmenting DNA and adding specialized adapters to both ends to create a sequencing library. In Illumina sequencing workflows, these adapters contain complementary sequences that allow DNA fragments to bind to the flow cell. Fragments can then be amplified and purified.

• Sequencing

During the sequencing step of the NGS workflow, the library is loaded into the flow cell and placed on the sequencer. Clusters of DNA fragments are amplified in a process called cluster generation, resulting in millions of copies of single-stranded DNA. On most Illumina sequencing instruments, clustering occurs automatically.

• Data Analysis

After sequencing, the instrument software identifies nucleotides (a process called base calls) and the predicted accuracy of those base calls. During data analysis, you can import sequencing data into standard analysis tools or build your own pipeline.

Advantages of NGS

• No prior knowledge of genome or genome characteristics is required
• It provides single nucleotide resolution and can detect related genes, alternative splicing transcripts, allele variations and single nucleotide polymorphisms
• Higher signal dynamic range
• Requires less DNA/RNA as input

Applications of NGS

• Rapid sequencing of the whole genome
• Deep sequencing target area
• Use RNA sequencing (RNA Seq) to discover new RNA variants and splicing sites, or quantify mRNA for gene expression analysis
• Analysis of epigenetic factors, such as genome wide DNA methylation and DNA protein interactions
• Sequencing cancer samples to study rare somatic variants, tumor subclones, etc
• Study human microbiome
• Identify new pathogens