Nucleic acid amplification technique

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List of terms related to Nucleic acid amplification technique

Editor-In-Chief: C. Michael Gibson, M.S., M.D. [1]

Overview

Nucleic acid amplification techniques (NAAT) are biochemistry and molecular biology methods that involve the in-vitro synthesis of many copies of DNA or RNA from one original template.

Types of Nucleic Acid Amplification Techniques (NAAT)

Ligase chain reaction

Main article: Ligase chain reaction

A ligase chain reaction is a DNA amplification technique based upon the ligation of oligonucleotide probes. The probes are designed to exactly match two adjacent sequences of a specific target DNA. The chain reaction is repeated in three steps in the presence of excess probe:

1. Heat denaturation of double-stranded DNA
2. Annealing of probes to target DNA
3. Joining of the probes by thermostable DNA ligase

After the reaction is repeated for 20-30 cycles, the production of ligated probe is measured.

Polymerase chain reaction

Main article: Polymerase chain reaction

Polymerase chain reaction (PCR) is an in vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include:

• Thermal denaturation of the double-stranded target molecules
• Annealing of the primers to their complementary sequences
• Extension of the annealed primers by enzymatic synthesis with DNA polymerase

The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.

Self-sustained sequence replication

Main article: Self-sustained sequence replication

Self-sustained sequence replication is an isothermal in-vitro nucleotide amplification process. The process involves the concomitant action of an RNA-directed DNA polymerase, ribonuclease(s), and DNA-directed RNA polymerases to synthesize large quantities of sequence-specific RNA and DNA molecules.