Application of Single-Tube Tri-Primer ARMS-PCR to Detect the NFKB1 ATTG Insertion/Deletion Polymorphism

Genetic Testing and Molecular Biomarkers
Jiashuo ChaoYan Deng

Abstract

The -94 ATTG insertion/deletion polymorphism (rs28362491) is an important functional polymorphism in the NFKB1 gene. It has been shown that rs28362491 is associated with many diseases. The purpose of this study was to establish a simple and reliable method to detect the ATTG insertion/deletion polymorphism. On the basis of the amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method, a single-tube tri-primer ARMS-PCR method was developed to detect the ATTG insertion/deletion polymorphism in 93 samples. The results of the single-tube tri-primer ARMS-PCR method were validated by DNA sequencing. After optimization of the PCR conditions, the single-tube tri-primer ARMS-PCR was established to detect the insertion/deletion polymorphism using agarose gel electrophoresis. In 93 volunteers, the genotype frequencies were 30.1% for Ins/Ins, 19.4% for Del/Del, and 50.5% for Ins/Del, respectively. The results of the single-tube tri-primer ARMS-PCR method were consistent with the results of DNA sequencing. This single-tube tri-primer ARMS-PCR is a reliable, simple, and cost-efficient genotyping method for the detection of the ATTG insertion/deletion polymorphism in the NFKB1 gene.

References

Aug 28, 2001·Nucleic Acids Research·S YeI N Day
Dec 22, 2006·Nature Reviews. Molecular Cell Biology·Neil D Perkins
Feb 7, 2015·Genetic Testing and Molecular Biomarkers·Hamzeh Mesrian TanhaSadeq Vallian
Apr 12, 2015·Journal of Virological Methods·Nathália DelvauxElisabeth Lampe
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Feb 3, 2016·Cancer Biomarkers : Section a of Disease Markers·Ebrahim Eskandari-NasabShadi Amininia

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Citations

Aug 24, 2019·Journal of Clinical Laboratory Analysis·Chenxi JinWeifeng Zhu
Mar 26, 2021·Frontiers in Genetics·Dongyong LvLiangwen Yu

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Methods Mentioned

BETA
chip
electrophoresis
genotyping
PCR
PCRs

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