Next Generation Sequencing

Next Generation Sequencing (NGS)

A massively parallel sequencing technology that allows for the sequencing of millions of DNA fragments simultaneously.

Sanger Sequencing

A traditional method of DNA sequencing that reads one DNA sequence at a time.

Throughput

The amount of DNA that can be sequenced at one time; NGS offers millions of simultaneous sequences compared to Sanger's single sequence.

Cluster Generation

The process of amplifying DNA fragments on a flow cell to create clusters of identical sequences for sequencing.

Sequencing by Synthesis (SBS)

The core method used in Illumina NGS to determine the nucleotide sequence by synthesizing complementary strands.

Whole Exome Sequencing (WES)

A method that sequences all protein-coding regions in the genome, targeting about 1-2% of the genomic content.

RNA-seq

A technique for studying the transcriptome to analyze gene expression levels and alternative splicing.

Library Construction

The first step in Illumina NGS, where DNA is fragmented and adapters are added for sequencing preparation.

Data Analysis

The final step in NGS where raw sequencing signals are translated into meaningful biological data.

Bridge PCR

A method during cluster generation where single-stranded DNA forms a bridge to amplify and create clusters on the flow cell.

Base Calling

The process of translating fluorescent signals into nucleotide sequences in NGS data analysis.

Target Enrichment

The initial step in WES where probes are used to capture specific exon sequences from fragmented DNA.

Quantification in RNA-seq

The measurement of gene expression levels based on the number of sequence reads mapped to specific genes.

Amplification

The process of increasing the number of copies of DNA fragments during cluster generation in NGS.

Fluorescently Labeled Nucleotide

Nucleotides used in SBS that emit fluorescence to indicate which base is being added during sequencing.

Alignment

The process of aligning sequenced reads to a reference genome to identify variants during data analysis.