Accurate Zygote-Specific Discrimination of Single-Nucleotide Polymorphisms Using Microfluidic Electrochemical DNA Melting Curves

Abstract Background: Several methods for detection of single nucleotide polymorphisms (SNPs; e.g., denaturing gradient gel electrophoresis and denaturing HPLC) are indirectly based on the principle of differential melting of heteroduplex DNA. We present a method for detecting SNPs that is directly based on this principle. Methods: We used a double-stranded DNA-specific fluorescent dye, SYBR Green I (SYBR) in an efficient system (PE 7700 Sequence Detector) in which DNA melting was controlled and monitored in a 96-well plate format. We measured the decrease in fluorescence intensity that accompanied DNA duplex denaturation, evaluating the effects of fragment length, dye concentration, DNA concentration, and sequence context using four naturally occurring polymorphisms (three SNPs and a single-base deletion/insertion). Results: DNA melting analysis (DM) was used successfully for variant detection, and we also discovered two previously unknown SNPs by this approach. Concentrations of DNA amplicons were readily monitored by SYBR fluorescence, and DNA amplicon concentrations were highly reproducible, with a CV of 2.6%. We readily detected differences in the melting temperature between homoduplex and heteroduplex fragments 15–167 bp in length and differing by only a single nucleotide substitution. Conclusions: The efficiency and sensitivity of DMA make it highly suitable for the large-scale detection of sequence variants.

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Development of a simple method for the determination of single nucleotide polymorphisms

Cell Biology International ◽

10.1042/cbi034s075c ◽

2010 ◽

Vol 34 (8) ◽

pp. S75-S75

Author(s):

Weifeng Zhu ◽

Zhuoqi Liu ◽

Daya Luo ◽

Xinyao Wu ◽

Fusheng Wan

Keyword(s):

Single Nucleotide Polymorphisms ◽

Nucleotide Polymorphisms ◽

Simple Method ◽

Single Nucleotide

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Single nucleotide polymorphisms in intron 5 of the feline myosin regulatory light chain gene detected by SSCP analysis

Animal Genetics ◽

10.1046/j.1365-2052.2000.00620.x ◽

2000 ◽

Vol 31 (4) ◽

pp. 281-282

Author(s):

A L Magnon ◽

K M Meurs ◽

M D Kittleson ◽

W A Ware

Keyword(s):

Single Nucleotide Polymorphisms ◽

Light Chain ◽

Regulatory Light Chain ◽

Sscp Analysis ◽

Chain Gene ◽

Myosin Regulatory Light Chain ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Light Chain Gene

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Single nucleotide polymorphisms of the factor IX gene for linkage analysis in the southern Chinese population

British Journal of Haematology ◽

10.1046/j.1365-2141.2000.02384.x ◽

2000 ◽

Vol 111 (2) ◽

pp. 540-543 ◽

Cited By ~ 2

Author(s):

Vivian Chan ◽

Irene Yam ◽

B. Yip ◽

P. Au ◽

M. K. Shing ◽

...

Keyword(s):

Single Nucleotide Polymorphisms ◽

Linkage Analysis ◽

Chinese Population ◽

Factor Ix ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Southern Chinese

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W14.373 A single nucleotide polymorphisms of the apolipoprotein A-V gene (SNP3, T-1131C) modulates postprandial lipoprotein metabolism

Atherosclerosis ◽

10.1016/s0021-9150(04)90372-3 ◽

2004 ◽

Vol 5 (1) ◽

pp. 86

Author(s):

C MARIN

Keyword(s):

Single Nucleotide Polymorphisms ◽

Lipoprotein Metabolism ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Apolipoprotein A ◽

V Gene ◽

Postprandial Lipoprotein

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Sex Differences in Childhood Associations between DNA Markers and General Cognitive Ability

Journal of Individual Differences ◽

10.1027/1614-0001.28.3.161 ◽

2007 ◽

Vol 28 (3) ◽

pp. 161-164 ◽

Cited By ~ 1

Author(s):

Rosalind Arden ◽

Nicole Harlaar ◽

Robert Plomin

Keyword(s):

Sex Differences ◽

Single Nucleotide Polymorphisms ◽

Cognitive Ability ◽

Dna Markers ◽

Community Sample ◽

Nucleotide Polymorphisms ◽

General Cognitive Ability ◽

Single Nucleotide ◽

The Difference

Abstract. An association between intelligence at age 7 and a set of five single-nucleotide polymorphisms (SNPs) has been identified and replicated. We used this composite SNP set to investigate whether the associations differ between boys and girls for general cognitive ability at ages 2, 3, 4, 7, 9, and 10 years. In a longitudinal community sample of British twins aged 2-10 (n > 4,000 individuals), we found that the SNP set is more strongly associated with intelligence in males than in females at ages 7, 9, and 10 and the difference is significant at 10. If this finding replicates in other studies, these results will constitute the first evidence of the same autosomal genes acting differently on intelligence in the two sexes.

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