Chapter 7, Problem 29P

Summary Introduction

To analyze:

In laboratory, the following dideoxy DNA sequencing gel is produced.

The double-stranded DNA sequence of this molecules to be determined.

The polarity of each strand is to be labeled.

Introduction:

DNA sequencing is the determination of a sequence of nucleotide bases (A, T, G, C) in a DNA.

Sanger sequencing is a method of sequencing a region of DNA that is up to $900$ base pairs in length. This process is also known as Chain termination method.

Sanger sequencing was first developed by Fred Sanger and his colleagues in $1977$.

This process involved the formation of many copies of the template DNA strand.

Dideoxynucleotides inhibit the chain elongation process of DNA polymerases. They are also abbreviated as $\text{2', 3'}$ Dideoxynucleotides (ddNTPs)- (ddATP, ddTTP, ddGTP, ddCTP).

The principle of Sanger sequencing is that sufficient time and material can produce at least one DNA sequence of every possible length with a tagged nucleotide at the end.

The tagged nucleotides get terminated because ddNTPs lack $3\text{'}-$ OH group that generally extends the chain.