Sanger Method DNA sequencing

Hello Dear All welcome to my channel. Here, In this tutorial we discuss the Sanger Method of DNA sequencing. The Sanger method, also known as chain-termination sequencing, is a technique used for DNA sequencing. In this method, the DNA template is copied in a reaction mixture containing deoxyribonucleotide triphosphates (dNTPs), a DNA polymerase enzyme, and a primer that binds to the template. The primer initiates the synthesis of a new DNA strand by adding complementary nucleotides to the template. In addition to dNTPs, the reaction mixture also contains small amounts of modified dNTPs, which lack a hydroxyl group at the 3' position. These modified dNTPs are called dideoxyribonucleotide triphosphates (ddNTPs) and are incorporated into the growing DNA strand by the polymerase enzyme, just like regular dNTPs. However, since ddNTPs lack the 3' hydroxyl group, they prevent the addition of further nucleotides to the growing DNA strand, thus terminating the chain. The reaction mixture is divided into four separate tubes, each containing a different ddNTP (ddATP, ddCTP, ddGTP, or ddTTP). As the polymerase enzyme adds nucleotides to the growing DNA strand, the reaction is terminated by the incorporation of a ddNTP, resulting in the production of DNA fragments of varying lengths that terminate at each nucleotide position. The fragments are separated by gel electrophoresis, which separates the fragments by size, with shorter fragments migrating further through the gel. The resulting gel is then exposed to X-ray film or a digital imaging system, which detects the labeled fragments, and the sequence of the original DNA template can be deduced by reading the sequence of the bands on the gel. The Sanger method is considered the gold standard for DNA sequencing and is still widely used in many laboratories today. However, it has largely been replaced by next-generation sequencing technologies, which are faster, more accurate, and able to sequence much larger amounts of DNA in a single run. Primer Designing lecture link: https://www.youtube.com/watch?v=jwOTap2mLFg Probe labelling : https://www.youtube.com/watch?v=unPVoOW8lhc #DNAsequening #sangermethod #biotechnology #csirnet #gate #rdnatechnology