Rapid single nucleotide polymorphism analysis by primer extension and capillary electrophoresis using polyvinyl pyrrolidone matrix

Barta, Csaba; Rónai, Zsolt; Sasvári-Székely, Mária; Guttman, András

doi:10.1002/1522-2683(200102)22:4<779::aid-elps779>3.0.co;2-4

Cited by 28 publications

(16 citation statements)

References 17 publications

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“…Although traditional submarine horizontal gel electrophoresis is still the method of choice in most instances (15 ), novel higher resolution and automated approaches have also been introduced recently. High-performance capillary gel electrophoresis is an instrumental approach with the capability of a single-base resolution (16,17 ). Electrophoretic microdevices were also demonstrated to be useful in high-throughput genotyping studies (18 -20 ).…”

confidence: 99%

Multicapillary Electrophoresis Analysis of Single-Nucleotide Sequence Variations in the Deoxycytidine Kinase Gene

et al. 2006

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Background: Investigation of the genetic background of complex traits is the focus of recent interest, as several common diseases or the individual response to treatments of various illnesses have not yet been explored. These studies require the development and implementation of reliable and large-scale genotyping methods. In this report, we introduce an efficient technique based on PCR-restriction fragment length sequence variation technique for the analysis of the ؊360CG and ؊201CT single-nucleotide sequence variations in the deoxycytidine kinase gene. Methods: A multicapillary gel electrophoresis instrument was used for the size determination of the generated DNA fragments. A healthy Hungarian population of 100 individuals was investigated to determine allele and genotype frequencies for the 2 sequence variations of interest. Results: We found that the occurrence of the minor allele is rather low, i.e., the frequency of both the ؊360G and ؊201T variants is 1%. Conclusions: Our technique can readily facilitate the analysis of these important sequence variations in other ethnic groups to clarify the role of these sequence variations in conjunction with arabinosylcytosine treatment in acute myeloid leukemia.

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confidence: 99%

Multicapillary Electrophoresis Analysis of Single-Nucleotide Sequence Variations in the Deoxycytidine Kinase Gene

et al. 2006

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“…A minisequencing technique with fluorescently labeled primers was used to detect the mutation at position 659 in intron 2 of the CYP21 gene, which causes a deficiency of the 21-hydroxylase enzyme [71]. Capillary electrophoresis was able to resolve single-stranded DNA in the range of 20-40 bases in 90 sec using poly (vinylpyrrolidone) as the separation matrix.…”

Section: Capillary Electrophoresismentioning

confidence: 99%

BioMEMS using electrophoresis for the analysis of genetic mutations

Thomas

Farquar

Sutton

et al. 2002

Expert Review of Molecular Diagnostics

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Biomedical microelectromechanical systems (BioMEMS) are rapidly emerging in many areas of genetic analysis. These devices demonstrate potential for rapid analysis using modular components capable of sample purification, amplification, mutation discrimination and detection on small, portable point-of-care instruments. Here, various approaches to genetic mutation detection and the modern analysis platform, capillary electrophoresis, will be briefly reviewed. Microfluidic devices will be discussed in relation to fabrication techniques, mutation detection using simple electrophoretic separations, multiplexed designs and modular functionalities, as well as challenges and issues surrounding this technology.

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“…Tomoka Nakazumi and Yusuke Hara were with Research Institute for Sustainable Chemistry, ISC, National Institute of Advanced Industrial Science and Technology, AIST, Central 5-2, 1-1-1 Higashi, Tsukuba 305-8565, Japan (e-mail: y-hara@ aist.go.jp). a fused silica capillary has been extensively investigated [5]- [10]. In this study, to prevent the EOF during the process of introducing the DNA samples, a non-ionic acrylamide polymer chain was adopted via chemical bonding to the fused silica wall.…”

Section: Introductionmentioning

confidence: 99%

Influence of the Sample Volume upon Injection on Capillary Gel Electrophoresis for the Separation of Small DNA Fragments

Nakazumi¹,

Hara²

2018

IJCEA

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In this study, we evaluated the influence of the sample volume (10 μL and 20 μL) upon the injection to a developed capillary gel electrophoresis (CGE) system designed using a short-length capillary (total length of 15 cm and effective length of 7.5 cm). The fused silica capillary was coated with an acrylamide linear polymer chain to prevent the electroosmotic flow in the direction opposite to the movement of DNA samples. The sieving polymer solution used was hydroxyethyl cellulose with a molecular weight of 1,300,000. Using this CGE system for the separation of a 100-bp DNA Ladder sample, the sample injection volume had a substantial effect on the electropherogram results. The error bars for migration time, mobility, and resolution length were much larger when using 20 μL of the sample than using 10 μL. The use of 10 μL of the sample provided highly reliable results. These findings indicated that developers of CGE equipment should pay close attention to the sample injection volume in order to measure small DNA samples with high accuracy.

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Rapid single nucleotide polymorphism analysis by primer extension and capillary electrophoresis using polyvinyl pyrrolidone matrix

Cited by 28 publications

References 17 publications

Multicapillary Electrophoresis Analysis of Single-Nucleotide Sequence Variations in the Deoxycytidine Kinase Gene

Multicapillary Electrophoresis Analysis of Single-Nucleotide Sequence Variations in the Deoxycytidine Kinase Gene

BioMEMS using electrophoresis for the analysis of genetic mutations

Influence of the Sample Volume upon Injection on Capillary Gel Electrophoresis for the Separation of Small DNA Fragments

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