Allele-specific competitive blocker PCR: a one-step method with applicability to pool screening

Human Mutation
A OrouH J Menzel

Abstract

We have developed a novel one-step pool screening PCR procedure which is based on the principles of amplification refractory mutation system (ARMS) and competitive oligonuleotide priming (COP) PCR. In addition to the usual primers, this approach uses two allele-specific competitive oligonucleotides, one of which is 3'-end labeled with a dideoxynucleotide and blocks amplification of the wild-type allele. An allele-specific product is generated only in the presence of the mutation. The introduction of an allele-specific competitive blocker oligonucleotide improves the specificity and robustness of ARMS-PCR. Further its sensitivity is dramatically increased, which allows detection of one mutant allele in a large excess of wild-type-bearing genomic DNA by electrophoresis in an ethidium bromide-stained agarose gel (up to 1 in 10(4) alleles). This makes the method ideal for nonradioactive pool screening. The successful application of the method has been demonstrated for four different point mutations, two in the apolipoprotein B gene (R3500Q, R3531C) which result in familial defective apolipoprotein B-100, one in the CFTR gene (R1162X), and one in the gene for lipoprotein lipase (G188E).

References

Nov 1, 1992·Trends in Genetics : TIG·L C Tsui
Apr 11, 1989·Nucleic Acids Research·R A GibbsC T Caskey

❮ Previous
Next ❯

Citations

Sep 28, 2001·Journal of Clinical Laboratory Analysis·A D HorvathV S Ganev
May 30, 2002·Mutation Research·Page B McKinzie, Barbara L Parsons
Dec 31, 1997·Mutation Research·B L Parsons, R H Heflich
May 2, 2014·Biomedical Microdevices·Scott O SundbergBruce K Gale
May 19, 2007·American Journal of Clinical Pathology·Angela Y C TanAlexander Dobrovic
Aug 27, 2010·Environmental and Molecular Mutagenesis·Barbara L ParsonsPage B McKinzie
Nov 28, 2012·International Journal of Cancer. Journal International Du Cancer·Jinyin ZhaoWeijun Chen
Jun 26, 2002·Carcinogenesis·Barbara L ParsonsRobert H Heflich
Jun 13, 2012·Chembiochem : a European Journal of Chemical Biology·Fumie TakeiKazuhiko Nakatani
Mar 11, 2018·Scientific Reports·Virginia Alvarez-GarciaNicholas R Leslie
Oct 17, 2006·Clinical Chemistry·Angela TanAlexander Dobrovic
May 19, 2001·Electrophoresis·J Drábek

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.