SangerVerfahren

The SangerVerfahren, also known as the Sanger method, is a chemical sequencing method for nucleic acids, such as DNA and RNA. This technique was developed in the 1970s by Frederick Sanger and his colleagues, and it remains a widely used method for sequencing and analyzing nucleic acids.

The SangerVerfahren works by using dideoxynucleotides (ddNTPs) to terminate DNA synthesis. In this method, a DNA polymerase is used to synthesize a DNA strand from a template, while ddNTPs are added to the reaction mixture. As the polymerase synthesizes the strand, the addition of ddNTPs leads to random termination of the strand at different points. This results in a mixture of DNA fragments of varying lengths.

The resulting fragments are then separated by size using electrophoresis, typically on a polyacrylamide gel. The fragments are detected using autoradiography, resulting in a pattern of bands that correspond to the sequence of the original DNA template. By comparing the pattern obtained from a DNA template with the sequence obtained from a known DNA sequence, researchers can deduce the sequence of the original DNA.

The SangerVerfahren is a relatively straightforward method for sequencing short DNA fragments. However, it has limitations in terms of speed, cost, and scalability for large-scale sequencing projects. It has largely been replaced by next-generation sequencing (NGS) technologies, which offer higher-throughput, faster, and more cost-effective sequencing methods. Nevertheless, the SangerVerfahren remains an important method in certain specialized applications and in teaching and training contexts.