What is allele specific oligonucleotide used for?
An allele-specific oligonucleotide (ASO) is a short piece of synthetic DNA complementary to the sequence of a variable target DNA. It acts as a probe for the presence of the target in a Southern blot assay or, more commonly, in the simpler Dot blot assay.
What is allele specific oligonucleotide hybridization?
Allele-specific oligonucleotide hybridization, or dot blotting, is a method for testing known mutations. Normal individuals show hybridization only with the normal probe; homozygous individuals show hybridization only with the mutant probe; and heterozygous individuals hybridize with both probes (normal and wild-type).
What is the principle of allele specific oligonucleotide?
Allele-specific oligonucleotides allow amplification of a sequence of DNA to which they are perfectly homologous (1), but not one containing a mismatch (2).
What is oligonucleotide ligation assay?
The oligonucleotide ligation assay (OLA) is a genotypic assay which has been used to identify point mutations in DNA for a variety of diseases (3, 5) and to detect drug resistance-associated mutations in HIV-1 subtype B (1, 7, 8, 15).
How does Heteroduplex analysis work?
Hetero-duplex analysis is a low-medium high-throughput SNP detection technique. During heteroduplex analysis, the target gene is PCR amplified and the amplified products are then denatured, re-annealed slowly to form four different double-stranded DNA molecules from the two alleles of the gene (Paniego et al., 2015).
What are SNP chips used for?
SNP chips are DNA microarrays that test genetic variation at many hundreds of thousands of specific locations across the genome. 5 They were initially designed for testing single nucleotide polymorphisms (SNPs) that are common in the population (>1 in 100 people).
How do you genotype a SNP?
SNP genotyping
- SNP genotyping is the measurement of genetic variations of single nucleotide polymorphisms (SNPs) between members of a species.
- In the first step, a genomic segment is amplified and attached to a bead through a PCR reaction with a biotinylated primer.
What is Heteroduplex PCR?
Heteroduplexes are PCR products that form as a result of cross-hybridization (i.e., recombination) between the competitor sequence and the experimental cDNA.