Automated Sanger Sequencing: Understanding the Steps

What are the correct steps for Automated Sanger Sequencing?

A. Denaturation, Primer annealing, Extension

B. Extension, Denaturation, Primer annealing

C. Primer annealing, Extension, Denaturation

D. Denaturation, Extension, Primer annealing

Answer:

The correct steps for Automated Sanger Sequencing are Denaturation, Primer Annealing, and Extension (A).

When delving into the realm of DNA sequencing, Automated Sanger Sequencing plays a crucial role in deciphering the genetic code. By understanding the intricate steps involved in this process, we can uncover the mysteries encoded within the DNA strands.

Exploring the Steps:

The first step in Automated Sanger Sequencing is Denaturation. This involves the heating of the DNA, which results in the separation of the two strands, creating single-stranded DNA molecules ready for further processing.

Following Denaturation, Primer Annealing takes place. During this step, specific primers bind to the single-stranded DNA, providing a starting point for the DNA polymerase to initiate synthesis.

Next comes Extension, where DNA polymerase adds nucleotides to the primer, elongating the DNA strand. The incorporation of dideoxynucleotides provides termination points at random intervals, crucial for later sequencing analysis.

Significance of Sequence Determination:

By following these meticulous steps, scientists can unravel the sequence of the DNA, unlocking valuable information about genetic traits, evolutionary relationships, and disease susceptibilities.

Moreover, the incorporation of dye-labeled dideoxynucleotides in Sanger Sequencing revolutionized the field, allowing for precise termination points and accurate sequencing results.

Legacy of Frederick Sanger:

Frederick Sanger's groundbreaking work in DNA sequencing earned him the prestigious Nobel Prize in Chemistry in 1980. His contributions to the field have paved the way for advancements in genomic research and personalized medicine.