Microplate array diagonal gel electrophoresis for mutation research in DNA banks

Day, Ian N.M.; Spanakis, Emmanuel; Chen, Xiaohe; O’Dell, Sandra D.

doi:10.1002/(sici)1522-2683(19990101)20:6<1250::aid-elps1250>3.0.co;2-5

Cited by 10 publications

(5 citation statements)

References 19 publications

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“…Products were resolved on a 7.5% polyacrylamide gel, 23 and genotypes were confirmed by two independent technicians blinded to subject outcome, with discrepancies resolved by repeat genotyping. The failure to genotype all individuals for all genotypes relates to quality and quantity of stored DNA and (being random) is not a source of confounding error.…”

Section: Genotypingmentioning

confidence: 99%

Genetic Variants of Angiotensin II Receptors and Cardiovascular Risk in Hypertension

et al. 2003

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Abstract-Renin-angiotensin systems may mediate cardiovascular disease pathogenesis through a balance of actions of angiotensin II on (potentially proatherogenic) constitutive type 1 (AT 1 R) and (potentially antiatherogenic) inducible type 2 (AT 2 R) receptors. We explored such potential roles in a prospective candidate gene association study. Cardiovascular end points (fatal, nonfatal, and silent myocardial infarction and coronary artery bypass surgery/angioplasty) were documented among 2579 healthy UK men (mean age, 56.1Ϯ3.5 years; median follow-up, 10.1 years) genotyped for the AT 1 R1166AϾC and the X chromosome located AT 2 R1675AϾG and 3123CϾA polymorphisms. Baseline characteristics, including blood pressure, were independent of genotype. The AT 1 R1166CC genotype was associated with relative cardiovascular risk (hazard ratio, 1.65 [1.05 to 2.59]; Pϭ0.03) independent of blood pressure. Systolic blood pressure was associated with risk (Pϭ0.0005), but this association was restricted to AT 2 R1675A allele carriers (PϽ0.00001), with G allele carriers protected from the risk associated with blood pressure (Pϭ0.18). Hypertensive carriers with the AT 2 R1675A/3123A haplotype were at most risk, with 37.5% having an event. This is the first study to demonstrate an association of AT 2 R genotype with coronary risk, an effect that was confined to hypertensive subjects and supports the concept that the inducible AT 2 R is protective. Conversely, the AT 1 R1166CC genotype was associated with cardiovascular risk irrespective of blood pressure. These data are important to our understanding of the divergent role of angiotensin II acting at its receptor subtypes and coronary disease pathogenesis and for the development of future cardiovascular therapies. Coronary vascular ACE drives Ang II synthesis, whose action on local AT 1 and inducible AT 2 receptors (AT 2 R) 2,3 may contribute to coronary heart disease (CHD) pathogenesis: AT 1 R activation causes vascular smooth muscle cell hypertrophy, extracellular matrix production, and local inflammation, driving atherogenesis and plaque rupture, 4 whereas AT 2 R agonism inhibits vascular cell proliferation 5 and may be antiatherogenic. 6 The balance between AT 1 R and AT 2 R activation may therefore influence CHD risk. However, this remains difficult to explore, and supportive data are sparse. Studies involving selective AT 1 R antagonism are perhaps less informative than they might at first appear: although lowering CHD risk more than equihypotensive ␤-blockade, 7 this may be partly mediated through AT 2 R agonism-loss of negative feedback raising Ang II levels and hence binding to the vacant AT 2 R. 8Could there be a role for both the AT 1 R and AT 2 R in the development of CHD? Genetic studies may provide insight. A polymorphism of the AT 1 R gene exists at position 1166, where the C (rather than A) allele is associated with increased Ang II responsiveness. 9 Meanwhile, the A (rather than G) allele at position 1675 of the X-chromosomal AT 2 R gene is associated with a greater ...

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Section: Genotypingmentioning

confidence: 99%

Genetic Variants of Angiotensin II Receptors and Cardiovascular Risk in Hypertension

et al. 2003

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“…Day et al (8,9) described temporal-thermalramp electrophoresis MADGE (melt MADGE), in which the analysis of duplex melting provides a method of de novo mutation scanning. Using a purpose-built temporal-thermal-ramp-electrophoresis apparatus for programmable melting display (PMD) (melt-MADGE), a DGGE-like analysis was undertaken in MADGE format.…”

Section: Temporal Thermal Ramp Electrophoresis Madge (Melt-madge)mentioning

confidence: 99%

Madge - Microplate Array Diagonal Gel Electrophoresis

Stanković¹,

Alavantić²,

Majkić-Singh³

2009

Journal of Medical Biochemistry

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Madge - Microplate Array Diagonal Gel ElectrophoresisMicroplate array diagonal gel electrophoresis (MADGE) was invented for molecular genetic epidemio-logical studies. MADGE is a highly flexible, cost effective, microplate compatible solution to high throughput electrophoresis needs. It enables several thousands to million gel lines per day for direct assay of single-base variation in different capacity laboratories. Variants of the standard 96-well MADGE include: 96-well stretch-MADGE, 192-well MADGE, 384-well MADGE, and 768-well MADGE. Melt-MADGE combines the temporal thermal ramp apparatus to achieve similar throughput for de novo mutation scanning. Basic MADGE principles and procedures, preparation of MADGE gels, electrophoresis, visualization and analysis of these gels, as well as modifications of the basic 96-well MADGE will be discussed in detail. For the first time in our country, this revolutionary polyacrylamid electrophoresis was done in 1998. We shortly review our studies which used MADGE for high throughput genotyping of the apolipoprotein E. MADGE and melt-MADGE will have an important role in the future genetic research of complex diseases and especially in pharmacogenomics.

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“…Melt-MADGE, a version that allows applying a temperature gradient during gel migration, was also shown. Melt-MADGE opens the path for further applications apart from SNP genotyping (Day et al 1999) such as SNP discovery and mutation detection. Table 4.1…”

Section: Gel-based Analysismentioning

confidence: 99%

An Overview of Genotyping and Single Nucleotide Polymorphisms (SNP)

Gut

2004

Molecular Analysis and Genome Discovery

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Microplate array diagonal gel electrophoresis for mutation research in DNA banks

Cited by 10 publications

References 19 publications

Genetic Variants of Angiotensin II Receptors and Cardiovascular Risk in Hypertension

Genetic Variants of Angiotensin II Receptors and Cardiovascular Risk in Hypertension

Madge - Microplate Array Diagonal Gel Electrophoresis

An Overview of Genotyping and Single Nucleotide Polymorphisms (SNP)

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