Sanger sequencing, also known as the chain termination method, is a foundational laboratory technique used to determine the exact nucleotide sequence of a DNA molecule. Developed by Frederick Sanger and his colleagues in 1977, it served as the gold standard method for DNA sequencing for nearly four decades and was the primary technology used to map the human genome during the Human Genome Project. The Core Principle: Chain Termination
The technique mimics natural DNA replication but introduces a clever “stop” mechanism using two types of nucleotides:
Deoxynucleotide triphosphates (dNTPs): Standard DNA building blocks (A, T, C, G) that allow the DNA strand to keep growing.
Dideoxynucleotide triphosphates (ddNTPs): Modified building blocks that lack a 3’-hydroxyl (OH) group. Without this chemical group, DNA polymerase cannot attach the next nucleotide, permanently stopping strand extension. The 4-Step Sequencing Workflow
How to Conduct Sanger Sequencing | Thermo Fisher Scientific – RU
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