Question

Draw a workflow of the original Sanger Sequencing and explain how the automated Sanger method is more efficient.

Answer 1

The Original Sanger Sequencing :

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.

In recent days, the higher capacity sequencing method is completely changed into the next-gen sequencing method.

It can generate the DNA sequence which is greater than 500 nucleotides.

In this method, the following is required:

DNA polymerase,

DNA primer

dNTP deoxynucleotide

ddNTPdideoxynucleotide

There is the formation of a phosphodiester bond between the two nucleotide

The automated danger method is more efficient in terms of providing large amount of DNA content. It can achieve the approximate read length of 800 Bp

• In order to study the structure of the genome structure
• Single nucleotide polymorphism (SNP) detection, is done using this method.
• heteroduplex analysis, single-strand conformation polymorphism (SSCP), and short tandem repeat (STR) analysis.
• Resolving DNA fragments according to differences in size and/or conformation is the most critical step in studying these features of the genome.

The below picture shows the Super Sanger Sequencing