Detection of Single-Nucleotide Polymorphisms Using an ON–OFF Switching of Regenerated Biosensor Based on a Locked Nucleic Acid-Integrated and Toehold-Mediated Strand Displacement Reaction

2014 ◽  
Vol 86 (5) ◽  
pp. 2543-2548 ◽  
Author(s):  
Zhong Feng Gao ◽  
Yu Ling ◽  
Lu Lu ◽  
Ning Yu Chen ◽  
Hong Qun Luo ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Single Nucleotide Polymorphisms ◽  
Locked Nucleic Acid ◽  
Displacement Reaction ◽  
Nucleotide Polymorphisms ◽  
Strand Displacement ◽  
Single Nucleotide ◽  
Strand Displacement Reaction

scholarly journals A Locked Nucleic Acid Probe Based on Selective Salt-Induced Effect Detects Single Nucleotide Polymorphisms

2015 ◽  
Vol 2015 ◽  
pp. 1-6
Author(s):  
Jing Zhang ◽  
Huizhe Wu ◽  
Qiuchen Chen ◽  
Pengfei Zhao ◽  
Haishan Zhao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Single Nucleotide Polymorphisms ◽  
Room Temperature ◽  
Genetic Mutation ◽  
Locked Nucleic Acid ◽  
Nucleotide Polymorphisms ◽  
Buffer Solution ◽  
Single Nucleotide ◽  
Real Time Quantitative Pcr ◽  
Pcr Products

Detection of single based genetic mutation by using oligonucleotide probes is one of the common methods of detecting single nucleotide polymorphisms at known loci. In this paper, we demonstrated a hybridization system which included a buffer solution that produced selective salt-induced effect and a locked nucleic acid modified 12 nt oligonucleotide probe. The hybridization system is suitable for hybridization under room temperature. By using magnetic nanoparticles as carriers for PCR products, the SNPs (MDR1C3435T/A) from 45 volunteers were analyzed, and the results were consistent with the results from pyrophosphoric acid sequencing. The method presented in this paper differs from the traditional method of using molecular beacons to detect SNPs in that it is suitable for research institutions lacking real-time quantitative PCR detecting systems, to detect PCR products at room temperature.

Locked nucleic acid (LNA) induced effect on the hybridization and fluorescence properties of oligodeoxyribonucleotides modified with nucleobase-functionalized DNA monomers

2015 ◽  
Vol 13 (26) ◽  
pp. 7236-7247 ◽  
Author(s):  
Mamta Kaura ◽  
Patrick J. Hrdlicka
Keyword(s):  
Nucleic Acid ◽  
Single Nucleotide Polymorphisms ◽  
Locked Nucleic Acid ◽  
Fluorescence Properties ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide

Mixmer oligonucleotides modified with LNA and C5-pyrene-functionalized DNA monomers are shown to display interesting fluorescence properties for the discrimination of single nucleotide polymorphisms (SNPs).

scholarly journals Homogeneous Real-Time Detection of Single-Nucleotide Polymorphisms by Strand Displacement Amplification on the BD ProbeTec ET System

2003 ◽  
Vol 49 (10) ◽  
pp. 1599-1607 ◽  
Author(s):  
Sha-Sha Wang ◽  
Keith Thornton ◽  
Andrew M Kuhn ◽  
James G Nadeau ◽  
Tobin J Hellyer
Keyword(s):  
Single Nucleotide Polymorphisms ◽  
Dna Sequencing ◽  
Real Time ◽  
Whole Blood ◽  
Nucleotide Polymorphisms ◽  
Strand Displacement ◽  
Strand Displacement Amplification ◽  
Single Nucleotide ◽  
Buccal Swabs ◽  
Real Time Detection

Abstract Background: The BD ProbeTec™ ET System is based on isothermal strand displacement amplification (SDA) of target nucleic acid coupled with homogeneous real-time detection using fluorescent probes. We have developed a novel, rapid method using this platform that incorporates a universal detection format for identification of single-nucleotide polymorphisms (SNPs) and other genotypic variations. Method: The system uses a common pair of fluorescent Detector Probes in conjunction with unlabeled allele-specific Adapter Primers and a universal buffer chemistry to permit analysis of multiple SNP loci under generic assay conditions. We used Detector Probes labeled with different dyes to facilitate differentiation of two alternative alleles in a single reaction with no postamplification manipulation. We analyzed six SNPs within the human β2-adrenergic receptor (β2AR) gene, using whole blood, buccal swabs, and urine samples, and compared results with those obtained by DNA sequencing. Results: Unprocessed whole blood was successfully genotyped with as little as 0.1–1 μL of sample per reaction. All six β2AR assays were able to accommodate ≥20 μL of unprocessed whole blood. For the 14 individuals tested, genotypes determined with the six β2AR assays agreed with DNA sequencing results. Conclusion: SDA-based allelic differentiation on the BD ProbeTec ET System can detect SNPs rapidly, using whole blood, buccal swabs, or urine.

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