ArticleMolecular genetics & genomic medicine2019
MTHFR C677T polymorphism analysis: A simple, effective restriction enzyme-based method improving previous protocols.
Article in Molecular genetics & genomic medicine, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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12 citing papers in PubMed.
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- Relationship between MTHFR gene polymorphism and risk of thrombosis in postoperative patients with colorectal cancer.Experimental and therapeutic medicine · 2023Article
- Down-regulation of COL1A1 inhibits tumor-associated fibroblast activation and mediates matrix remodeling in the tumor microenvironment of breast cancer.Open life sciences · 2023Article
- Prevalence of polycystic kidney disease in Persian and Persian-related cats in western Mexico.Journal of feline medicine and surgery · 2022Article
- MTHFR Gene-Polymorphism and Infertile Men in Indian Population: A Systematic Literature Review.Cureus · 2022Review
- The transcriptome profile of human trisomy 21 blood cells.Human genomics · 2021Article
- Correlation of methylation status in MTHFR promoter region with recurrent pregnancy loss.Journal, genetic engineering & biotechnology · 2021Article
- One-carbon pathway and cognitive skills in children with Down syndrome.Scientific reports · 2021Article
- A floating thrombus detected by CT in the descending aorta in an obese patient: A possible genetic-environmental interaction.Radiology case reports · 2021Article
- MTHFR C677T polymorphism analysis: A simple, effective restriction enzyme-based method improving previous protocols.Molecular genetics & genomic medicine · 2019Article
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Abstract
background5,10-Methylentetrahydrofolate reductase (MTHFR) C677T polymorphism is one of the most studied genetic variations in the human genome. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is one of the most used techniques to characterize the point mutations in genomic sequences because of its suitability and low cost. The most widely used method for the MTHFR C677T polymorphism characterization was developed by Frosst et al. (1995) but appears to have some technical limitations. The aim of this study was to propose a novel PCR-RFLP method for the detection of this polymorphism.
methodsIn order to retrieve all published articles possibly describing any PCR-RFLP methods useful to analyze MTHFR C677T polymorphism, we performed systematic queries on PubMed, using a combination of Boolean operators (AND/OR) and MeSH terms. Amplify software was used in order to design a new primer pair following the optimal standard criteria. Primer-BLAST software was used to check primer pair's biological specificity.
resultsThe analysis of previous literature showed that PCR-RFLP method remains the most used technique. None of the 108 primer pairs described was ideal with regard to main accepted primer pair biochemical technical parameters. The new primer pair amplifies a DNA-fragment of 513 base pair (bp) that, in the presence of the polymorphism, is cut by Hinf I enzyme in two pieces of 146 bp and 367 bp and clearly visible on 2% agarose gel. The level of expertise and the materials required are minimal and the protocol takes one day to carry out.
conclusionOur original PCR-RFLP strategy, specifically designed to make the analysis optimal with respect to PCR primers and gel analysis, fits the ideal criteria compared to the widely used strategy by Frosst et al (1995) as well as any other PCR-RFLP strategies proposed for MTHFR C677T polymorphism genotyping to date.
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