DNA sequencing is a powerful technique that allows scientists to decipher the genetic code of an organism. One method of DNA sequencing is the Sanger sequencing method, which revolutionized the field of molecular biology.

Category : DNA Sequencing | Sub Category : Sanger Sequencing Method Posted on 2024-02-07 21:24:53

DNA sequencing is a powerful technique that allows scientists to decipher the genetic code of an organism. One method of DNA sequencing is the Sanger sequencing method, which revolutionized the field of molecular biology.

Developed by Frederick Sanger in the 1970s, Sanger sequencing is a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication. This method allows for the determination of the sequence of nucleotides in a DNA molecule.

The Sanger sequencing process begins with the preparation of a DNA sample to be sequenced. The DNA is denatured into single strands and then annealed with a short DNA primer that is complementary to a known sequence adjacent to the region of interest. DNA polymerase is then used to synthesize a new DNA strand complementary to the template DNA, incorporating both normal deoxynucleotides and chain-terminating dideoxynucleotides.

The chain-terminating dideoxynucleotides lack a 3' hydroxyl group, preventing the addition of further nucleotides and terminating DNA synthesis. This results in a series of DNA fragments of varying lengths that correspond to the position of each chain-terminating nucleotide. The DNA fragments are then separated by size using gel electrophoresis, allowing for the determination of the DNA sequence.

Sanger sequencing revolutionized the field of molecular biology by allowing scientists to rapidly and accurately determine the sequence of DNA molecules. This method has been used in numerous research studies, clinical diagnostics, and forensics applications.

In recent years, newer high-throughput sequencing technologies, such as next-generation sequencing, have largely replaced Sanger sequencing for large-scale genome sequencing projects due to their increased speed and lower cost. However, Sanger sequencing remains a valuable tool for sequencing small DNA fragments with high accuracy and reliability.

Overall, the Sanger sequencing method has had a profound impact on the field of molecular biology and continues to be used in various applications, contributing to our understanding of the genetic code and driving advancements in scientific research and medical diagnostics.