Master the Basics of DNA Sequencing

DNA Sequencing Steps

• The first step in DNA sequencing is to prepare the template by denaturing the DNA and adding a primer.
• The second step in DNA sequencing is to add DNA polymerase and nucleotides to the template, allowing the synthesis of new DNA strands to begin.

Nucleotides in DNA Sequencing

• ddNTPs (dideoxynucleotides) are used to terminate DNA synthesis in a base-specific manner, allowing for the determination of the DNA sequence.
• dNTPs (deoxynucleotides) are used as building blocks for the synthesis of new DNA strands during DNA sequencing.

Base Selective Termination

• During base selective termination of DNA synthesis, the incorporation of a ddNTP into the growing DNA strand causes the synthesis reaction to terminate, resulting in a set of DNA fragments of varying lengths that are specific to each base.

Capillary Electrophoresis

• Capillary electrophoresis is a technique used in DNA sequencing to separate and analyze the DNA fragments generated during the sequencing reaction, based on their size and charge.

Sequencing Reaction

• The two reactions that occur simultaneously during the sequencing reaction are the extension of the primer and the termination of the synthesis reaction, allowing for the simultaneous synthesis and termination of DNA strands.