Single strand conformation polymorphism (SSCP) as a quick and reliable method to genotype M235T polymorphism of angiotensinogen gene

Bettinaglio, Paola; Galbusera, Andrea; Caprioli, Jessica; Orisio, Silvia; Perna, Annalisa; Arnoldi, Federica; Bucchioni, Sara; Noris, Marina

doi:10.1016/s0009-9120(02)00308-9

Cited by 12 publications

(8 citation statements)

References 19 publications

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“…In the case of PCR-ASO, the signal-to-noise ratio of the hybridization of the nucleotides to the PCR product might be a limiting factor. For SSCP, strict control of the temperature of the gel during separation is of extreme importance (10). Together, these facts suggest that the improved PCR-RFLP protocol presented here could serve as a widely available method with enhanced accuracy for AGT M235T polymorphism determination.…”

Section: Discussionmentioning

confidence: 84%

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Mistyping of Angiotensinogen M235T Alleles

et al. 2006

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Section: Discussionmentioning

confidence: 84%

“…There are alternatives to the use of PCR-RFLP for determining the AGT M235T polymorphism, such as direct sequencing, PCR-ASO (8), MS-PCR (9) and single strand conformation polymorphism (SSCP) (10). However, these methods require special instruments and personnel, which may well not be available in most clinics.…”

Section: Discussionmentioning

confidence: 99%

Mistyping of Angiotensinogen M235T Alleles

et al. 2006

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“…Genomic DNA was extracted from peripheral blood leukocytes using standard procedures (Promega genomic DNA purification kit, USA). The Kir6.1 gene was amplified by PCR and screened using the PCR-SSCP method (17,18). Primers of the Kir6.1 gene were designed using Primer3 (http://frodo.wi.mit.edu/primer3/) ( Table I).…”

Section: Methodsmentioning

confidence: 99%

Mutational analysis of the Kir6.1 gene in Chinese hypertensive patients treated with the novel ATP-sensitive potassium channel opener iptakalim

Duan

Cui

Wang

2011

Experimental and Therapeutic Medicine

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Abstract. Iptakalim is a novel K ATP opener with antihypertensive properties. The Kir6.1 gene is one of the candidate genes that may influence the response to iptakalim in hypertensive patients. We aimed to ascertain whether a mutation in the coding region of the Kir6.1 gene is present in Chinese Han hypertensive patients. The study population included 162 Chinese Han hypertensive patients (81 men and 81 women with a mean age of 55±9 years). Mutational analysis of the coding region of Kir6.1 was performed using PCR-SSCP and direct sequencing. No missense or nonsense mutations were found in these samples, while a single base pair substitution (C to T) at nucleotide position 111 in exon 2 of the coding region was noted in four patients (one female and three male), who were all heterozygous for the mutation. This C to T substitution did not result in an amino acid substitution (Ile37Ile, silent mutation). In the remaining 158 patients, no mutation was detected. The blood pressure of these four patients carrying the I37I polymorphism was well controlled by iptakalim. No mutation that alters the primary structure of Kir6.1 was detected in Chinese Han hypertensive patients. The results indicate that abnormality in the primary structure of Kir6.1 is not involved in the genetic pathogenesis of essential hypertension in Chinese Han hypertensive patients and has no effect on the BP response to iptakalim treatment.

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“…• antisense: 5'-GCCAGAGCCAGCAGAGAGGTTTG-3' 25 The amplified fragments underwent sequence analysis in the MegaBace Analyzer (Amersham Biosciences), using Dynamic ET Dye Terminator cycle sequencing reagent; at the end of the run, the chromatogram was analyzed in the Sequence Analyzer (Figure 1). …”

Section: Genetic Analysismentioning

confidence: 99%