Allele-specific oligonucleotide

Allele-Specific Oligonucleotide (ASO) is a technique used in genetic testing to detect the presence of specific alleles in a DNA sample. ASO probes are short, single-stranded DNA sequences that are designed to hybridize with a complementary DNA or RNA sequence in a sample. The technique is used in a variety of applications, including genetic diagnosis, forensic science, and molecular biology research.

Overview

Allele-specific oligonucleotide probes are designed to be complementary to a specific DNA or RNA sequence. The probes are labeled with a radioactive or fluorescent marker, which allows for the detection of hybridization. The technique is highly specific, as the probes will only hybridize with a perfectly complementary sequence. This allows for the detection of specific alleles, even in a complex mixture of DNA or RNA.

Applications

ASO probes are used in a variety of applications. In genetic diagnosis, they can be used to detect the presence of specific mutations or polymorphisms that are associated with a particular disease. In forensic science, they can be used to identify individuals based on their unique DNA profile. In molecular biology research, they can be used to study gene expression and regulation.

Procedure

The procedure for using ASO probes involves several steps. First, the DNA or RNA sample is denatured to separate the two strands. The ASO probe is then added to the sample, and the mixture is heated to a temperature that allows for hybridization. The probe will bind to any complementary sequences in the sample. The sample is then washed to remove any unbound probe, and the bound probe is detected using a radioactive or fluorescent marker.

Limitations

While ASO probes are highly specific, they do have some limitations. The technique requires a priori knowledge of the sequence to be detected, which means it cannot be used to discover new mutations or polymorphisms. In addition, the technique can be time-consuming and requires a high level of technical skill.

See Also

• Polymerase chain reaction
• DNA sequencing
• Genetic testing