A bead-based single nucleotide polymorphism (SNP) detection using melting temperature on a microchip

Kao, Pei-Chun; Ding, Shih‐Torng; Lin, Eric S.; Li, Kan-Chien; Wang, Lon; Lu, Yen‐Wen

doi:10.1007/s10404-014-1331-0

Cited by 8 publications

(11 citation statements)

References 44 publications

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“…It was aligned to the region of heater/thermometer on the ITO glass and bonded together by partially-curing bonding method. 13 Fig. 2(a) showed the prototype of our droplet-based device with electrical and uidic connections.…”

Section: Fabrication Of Microuidic Devicementioning

confidence: 99%

“…microbeads), the mobile microbeads have their surfaces immobilized with DNA targets or probes, and then being genotyped either statically or dynamically. [13][14][15] However, the solidphase-based platform is limited in reaction rate by diffusion kinetics and requires surface modication for extensive reagent immobilization to ensure effective binding for detection. In contrast, the liquid phase carries the sample in a bulk ow and is introduced for downstream reaction and detection.…”

Section: Introductionmentioning

confidence: 99%

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Automated melting curve analysis in droplet microfluidics for single nucleotide polymorphisms (SNP) genotyping

et al. 2017

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Section: Fabrication Of Microuidic Devicementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Automated melting curve analysis in droplet microfluidics for single nucleotide polymorphisms (SNP) genotyping

et al. 2017

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“…Rice molecular breeding can increase the efficiency of phenotypic selection, expand the range of available alleles, overcome the shortcomings of original varieties, shorten the breeding cycle and improve rice quality. At present, traditional methods for mutants identification, such as TaqMan probe, sequencing, gene chip and mass-spectrometry analysis are complicated to operate, with high cost and fail to detect unknown mutations [2]. Sanger sequencing is the golden standard for short DNA fragment analysis of the mutants.…”

Section: Introductionmentioning

confidence: 99%

A microfluidic system integrated one-step PCR and high-resolution melting analysis for rapid rice mutant detection

Zhu

et al. 2019

Biotechnology & Biotechnological Equipment

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Rice is a crucial crop to meet global food demands, therefore, it is necessary to develop rapid, efficient and inexpensive methods for screening mutants of important genes in molecular breeding. In this research, a simple microfluidic chip-integrated one-step polymerase chain reaction (PCR) and high-resolution melting (HRM) analysis were designed for the rapid screening of rice mutants. In order to complete the above processes, we also installed a self-developed functional instrument integrated with a temperature control system and a fluorescence detection department. This microfluidic platform was validated by analysing genes related to insect resistance, phenotype and yield of two important varieties of rice, Oryza sativa L. ssp. japonica. and Oryza sativa L. ssp. indica (93-11). The results showed that the platform could detect singlenucleotide mutations between the wild and the mutant rice plants in 1 h and the accuracy was consistent with gene sequencing regarded as the golden standard of the mutants screening. Compared to the traditional method (requiring approximately 3 h), this platform can distinguish mutants from wild types more rapidly. This system is a simple, integrated, disposable and lowcost tool for high-throughput screening of mutants in molecular breeding. More important, this portable instrument can be used as a point-of-care platform for nucleic acid diagnostics in many scientific fields. ARTICLE HISTORY

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“…It was then aligned to the region of the embedded heater and thermometers, and bonded onto the ITO glass using the partially cured bonding method. 34 The microchip then could be obtained as shown in Fig. 3.…”

Section: Fabricationmentioning

confidence: 99%

“…The use of microbeads in the continuous-flow temperature gradient can bring the advantage in multiplex capability, compared to complicated procedures required in this aspect in our previous work. 34 Our devices presented in this article will allow one type of DNA samples immobilized onto one type of microbeads upstream first. Various types of the microbeads can carry different DNA sequences-while passing through the temperature from 60 C to 85 C to conduct individual melting analysis on each microbead in serial, so multiplex, high-throughput capability is achieved.…”

Section: -11mentioning

confidence: 99%

Melting analysis on microbeads in rapid temperature-gradient inside microchannels for single nucleotide polymorphisms detection

Ding

Lin

et al. 2014

Biomicrofluidics

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A continuous-flow microchip with a temperature gradient in microchannels was utilized to demonstrate spatial melting analysis on microbeads for clinical Single Nucleotide Polymorphisms (SNPs) genotyping on animal genomic DNA. The chip had embedded heaters and thermometers, which created a rapid and yet stable temperature gradient between 60 C and 85 C in a short distance as the detection region. The microbeads, which served as mobile supports carrying the target DNA and fluorescent dye, were transported across the temperature gradient. As the surrounding temperature increased, the fluorescence signals of the microbeads decayed with this relationship being acquired as the melting curve. Fast DNA denaturation, as a result of the improved heat transfer and thermal stability due to scaling, was also confirmed. Further, each individual microbead could potentially bear different sequences and pass through the detection region, one by one, for a series of melting analysis, with multiplex, high-throughput capability being possible. A prototype was tested with target DNA samples in different genotypes (i.e., wild and mutant types) with a SNP location from Landrace sows. The melting temperatures were obtained and compared to the ones using a traditional tube-based approach. The results showed similar levels of SNP discrimination, validating our proposed technique for scanning homozygotes and heterozygotes to distinguish single base changes for disease research, drug development, medical diagnostics, agriculture, and animal production. V C 2014 AIP Publishing LLC.

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A bead-based single nucleotide polymorphism (SNP) detection using melting temperature on a microchip

Cited by 8 publications

References 44 publications

Automated melting curve analysis in droplet microfluidics for single nucleotide polymorphisms (SNP) genotyping

Automated melting curve analysis in droplet microfluidics for single nucleotide polymorphisms (SNP) genotyping

A microfluidic system integrated one-step PCR and high-resolution melting analysis for rapid rice mutant detection

Melting analysis on microbeads in rapid temperature-gradient inside microchannels for single nucleotide polymorphisms detection

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