FDA Approves Revuforj® as First Menin Inhibitor for Acute Leukemia with KMT2A Translocation

CHMP Recommends Approval of Opdivo® Plus Yervoy® for First-Line Treatment of MSI-H/dMMR Metastatic Colorectal Cancer

ADDF Invests $1.18M to Standardize p-tau217 Blood Tests for Alzheimer’s Diagnosis

GSK Reports Positive Interim Results for DREAMM-7 Trial of Blenrep in Multiple Myeloma

Molbio Diagnostics Invests $30 Million in OptraSCAN to Revolutionize Digital Pathology

BioNTech Acquires Chinese Oncology Firm Biotheus for $950M

The Care and Protection of Children Trust

CPCT Launches Groundbreaking Initiative to Provide Lifelong Free Treatment for Children with Spinal Muscular Atrophy (SMA) in Karnataka

Early Comprehensive Genomic Profiling Improves Cancer Treatment Outcomes, Study Shows

EMA Reverses Decision, Recommends Limited Approval for Eisai and Biogen’s Alzheimer’s Drug Leqembi

Exact Sciences Unveils Advances in Multi-Cancer Early Detection with Cancerguard™ Test

MSD and LaNova Medicines Enter Exclusive Global License Agreement for Novel Cancer Therapy, LM-299

Promising CAR T Cell Therapies from BMS and Cabaletta Bio Show Potential for Durable Remission in Severe Lupus

FDA Grants Accelerated Approval to PTC Therapeutics' KEBILIDI™, the First Gene Therapy for AADC Deficiency in the U.S.

Association for Molecular Pathology Publishes Joint Consensus Report on SARS-CoV-2 Genomic Surveillance for Hospitals and Public Health Laboratories

Philips India Unveils Advanced Azurion Imaging System to Revolutionize Cardiac and Neurovascular Care

Home
/
insight
/
details

The Enduring Legacy of Sanger Sequencing in Genomic Research

NextEdge Admin

01 May 2024

3 min 38 sec

Sanger sequencing, named after its developer Frederick Sanger, is a method used to determine the precise sequence of nucleotides in a DNA molecule. It was first introduced in 1977 and played a pivotal role in the early stages of genomics research, including the Human Genome Project.

The principle of Sanger sequencing involves DNA replication in vitro using DNA polymerase, with the addition of chain-terminating dideoxynucleotides (ddNTPs). These ddNTPs lack the 3′-OH group necessary for DNA chain elongation, causing DNA synthesis to halt when they are incorporated into the growing DNA strand. Each ddNTP is labeled with a fluorescent or radioactive tag corresponding to one of the four DNA bases (A, T, C, G).

After the sequencing reaction, the resulting DNA fragments are separated by size using gel electrophoresis. As the fragments migrate through the gel, their sizes are detected by fluorescence or autoradiography, revealing the sequence of the DNA template based on the order of termination.

In particular, Sanger sequencing found its niche in small-scale sequencing tasks such as single-gene studies, routine sequencing for cloning and genotype checking, and specialized projects. Unlike NGS, which excels in parallel sequencing and high-throughput applications, Sanger’s superiority in accuracy, cost-effectiveness, and speed proved advantageous in these contexts.

Moreover, Sanger sequencing complemented NGS in various scenarios. It filled gaps in NGS data in challenging regions with low coverage or difficult sequences, validated NGS results by re-sequencing critical sections of the genome and served as a benchmark for validating new NGS approaches.

For instance, in analyzing regions like the CEBPA gene, notorious for technical challenges including high GC content and repeat regions, NGS often produced suboptimal results, necessitating Sanger sequencing to ensure comprehensive mutation detection. Similarly, re-sequencing critical areas confirmed NGS findings, enhancing confidence in mutation calls. By conducting side-by-side comparisons, Sanger sequencing served as a reference for validating the accuracy of NGS methodologies before their integration into routine workflows.

Sanger sequencing’s enduring relevance in the genomic landscape is evident. Its combination with NGS technologies not only fills technical gaps but also enhances the accuracy and reliability of genomic data. Thus, it remains an indispensable tool for researchers and molecular laboratories and may continue to be so for decades to come.