scholarly journals High Resolution Melting (HRM) analysis on VviDXS to reveal muscats or non-muscats among autochthonous Greek wine producing grape varieties

OENO One ◽  
2016 ◽  
Vol 50 (3) ◽  
Author(s):  
Georgios Merkouropoulos ◽  
Ioannis Ganopoulos ◽  
Andreas Doulis ◽  
Nikolaos Nikolaou ◽  
Photini Mylona
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Single Step ◽  
Maturity Stage ◽  
Single Nucleotide ◽  
Breeding Programs ◽  
Hrm Analysis ◽  
Fruit Maturity ◽  
Grape Varieties ◽  
Grape Breeding

<p style="text-align: justify;">Muscat flavor in grapes is associated with a substitution of a Single Nucleotide Polymorphism (SNP) located at position 1822 (SNP<sub>1822G&gt;T</sub>) within the coding sequence of the <em>VviDXS</em> gene. Various methods, including the use of High Resolution Melting (HRM) analysis, have been suggested to discriminate different SNP allelic states including the molecular discrimination of the muscat from the non-muscat grape varieties, thus providing the ability to minimize lengthy grape breeding programs when selecting for grape muscat flavor before the fruit maturity stage. HRM analysis on the SNP<sub>1822</sub> was performed on a group of 128 wine producing grape varieties in order to separate the muscat from the non-muscat genotypes before they are used for further breeding activities. This approach could be used either as a single-step prescreening method or in accordance with recently published methodology to elucidate on varietal characterization and authentication as these are important requirements concerning nurseries, growers and winemakers.</p>

scholarly journals High-Resolution Melting Analysis of the spa Repeat Region of Staphylococcus aureus

2008 ◽  
Vol 54 (2) ◽  
pp. 432-436 ◽  
Author(s):  
Alex J Stephens ◽  
John Inman-Bamber ◽  
Philip M Giffard ◽  
Flavia Huygens
Keyword(s):  
Staphylococcus Aureus ◽  
High Resolution ◽  
High Resolution Melting ◽  
Single Step ◽  
Staphylococcal Protein A ◽  
Step Method ◽  
Hrm Analysis ◽  
Closed Tube ◽  
Spa Types ◽  
Blind Trials

Abstract Background: The staphylococcal protein A (spa) locus of Staphylococcus aureus contains a complex repeat structure and is commonly used for single-locus sequence-based genotyping. The real-time PCR platform supports genotyping methods that are single step and closed tube and potentially can be carried out simultaneously with diagnosis. We describe here a method for genotyping S. aureus using high-resolution melting (HRM) analysis of the spa polymorphic region X. Methods: The conventional PCR spa assay was modified and optimized for the Rotor-Gene 6000 instrument (Corbett Life Science). HRM analysis on the Corbett Rotor-Gene 6000 instrument was used to test 22 known spa sequences obtained from 44 diverse methicillin-resistant S. aureus (MRSA) isolates. Criteria for calling pairs of melting curves “same” or “different” were developed empirically by converting the data to difference graph format with one curve defined as the control. HRM curve comparison between runs was done to determine the portability of the method. The assay performance was assessed by genotyping uncharacterized isolates, carrying out blind trials, and comparing HRM profiles from different runs. Results: HRM analysis of 44 diverse MRSA isolates generated 20 profiles from 22 spa sequence types. The 2 unresolved HRM spa types differed by only 1 bp. Two blind trials demonstrated complete reproducibility with respect to calling the different spa types. Interrun comparisons of HRM curves were successfully developed, indicating the robustness of the method. Conclusion: Analysis of the spa locus by HRM resolves spa sequence variants. This single- and closed-tube single-step method for S. aureus genotyping can be easily combined with the interrogation of other genetic markers.

scholarly journals Differentiation of Mitragyna speciosa, a narcotic plant, from allied Mitragyna species using DNA barcoding-high-resolution melting (Bar-HRM) analysis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chayapol Tungphatthong ◽  
Santhosh Kumar J. Urumarudappa ◽  
Supita Awachai ◽  
Thongchai Sooksawate ◽  
Suchada Sukrong
Keyword(s):  
High Resolution ◽  
Dna Barcoding ◽  
High Resolution Melting ◽  
Dna Barcode ◽  
Herbal Medicines ◽  
Efficient Tool ◽  
Hrm Analysis ◽  
Mitragyna Speciosa ◽  
Leaf Powder

AbstractMitragyna speciosa (Korth.) Havil. [MS], or “kratom” in Thai, is the only narcotic species among the four species of Mitragyna in Thailand, which also include Mitragyna diversifolia (Wall. ex G. Don) Havil. [MD], Mitragyna hirsuta Havil. [MH], and Mitragyna rotundifolia (Roxb.) O. Kuntze [MR]. M. speciosa is a tropical tree belonging to the Rubiaceae family and has been prohibited by law in Thailand. However, it has been extensively covered in national and international news, as its abuse has become more popular. M. speciosa is a narcotic plant and has been used as an opium substitute and traditionally used for the treatment of chronic pain and various illnesses. Due to morphological disparities in the genus, the identification of plants in various forms, including fresh leaves, dried leaf powder, and finished products, is difficult. In this study, DNA barcoding combined with high-resolution melting (Bar-HRM) analysis was performed to differentiate M. speciosa from allied Mitragyna and to assess the capability of Bar-HRM assays to identify M. speciosa in suspected kratom or M. speciosa-containing samples. Bar-HRM analysis of PCR amplicons was based on the ITS2, rbcL, trnH-psbA, and matK DNA barcode regions. The melting profiles of ITS2 amplicons were clearly distinct, which enabled the authentication and differentiation of Mitragyna species from allied species. This study reveals that DNA barcoding coupled with HRM is an efficient tool with which to identify M. speciosa and M. speciosa-containing samples and ensure the safety and quality of traditional Thai herbal medicines.

scholarly journals A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 199
Author(s):  
Xiaochun Zhang ◽  
Huan Yu ◽  
Qi Yang ◽  
Ziwei Wang ◽  
Ruocheng Xia ◽  
...  
Keyword(s):  
High Resolution ◽  
Dna Barcoding ◽  
High Resolution Melting ◽  
Its Region ◽  
Its Sequencing ◽  
Melting Temperatures ◽  
Hrm Analysis ◽  
National Drug ◽  
Species Specific ◽  
Its1 And Its2

In recent years, trafficking and abuse of hallucinogenic mushrooms have become a serious social problem. It is therefore imperative to identify hallucinogenic mushrooms of the genus Psilocybe for national drug control legislation. An internal transcribed spacer (ITS) is a DNA barcoding tool utilized for species identification. Many methods have been used to discriminate the ITS region, but they are often limited by having a low resolution. In this study, we sought to analyze the ITS and its fragments, ITS1 and ITS2, by using high-resolution melting (HRM) analysis, which is a rapid and sensitive method for evaluating sequence variation within PCR amplicons. The ITS HRM assay was tested for specificity, reproducibility, sensitivity, and the capacity to analyze mixture samples. It was shown that the melting temperatures of the ITS, ITS1, and ITS2 of Psilocybe cubensis were 83.72 ± 0.01, 80.98 ± 0.06, and 83.46 ± 0.08 °C, and for other species, we also obtained species-specific results. Finally, we performed ITS sequencing to validate the presumptive taxonomic identity of our samples, and the sequencing output significantly supported our HRM data. Taken together, these results indicate that the HRM method can quickly distinguish the DNA barcoding of Psilocybe cubensis and other fungi, which can be utilized for drug trafficking cases and forensic science.

scholarly journals Genotyping of GATA4 Gene Variant (G296S) in Malaysian Congenital Heart Disease Subjects by Real-Time PCR High Resolution Melting Analysis

2013 ◽  
Vol 32 (2) ◽  
pp. 152-157
Author(s):  
Nora Fawzi ◽  
Ramachandran Vasudevan ◽  
Patimah Ismail ◽  
Mazeni Alwi ◽  
Ahmad Fazli Abdul Aziz ◽  
...  
Keyword(s):  
Congenital Heart Disease ◽  
Heart Disease ◽  
High Resolution ◽  
Congenital Heart ◽  
High Resolution Melting ◽  
New Technology ◽  
Cost Effective ◽  
Study Cohort ◽  
Hrm Analysis ◽  
Melting Analysis

Summary Background: Congenital heart disease (CHD) is the most common birth defect; however, the underlying etiology is unrecognized in the majority of cases. GATA-binding protein 4 (GATA4), a cardiac transcription factor gene, has a crucial role in the cardiogenesis process; hence, a number of heterozygote sequence variations were identified as a cause of CHD. G296S heterozygote variant is the most frequently reported GATA4 gene sequence alteration. This study aims to investigate the role of G296S variant of the GATA4 gene in Malaysian CHD subjects. Methods: We have investigated 86 Malaysian CHD subjects with cardiac septation defects for the presence of the GATA4 gene heterozygote variant (G296S) by the new technology of high resolution melting (HRM) analysis. Results: Genotyping of G296S (c.886G>A) by HRM analysis shows that all the sample genotypes were of the wild GG type genotype and the heterozygote mutant GA genotype was totally absent from this study cohort. Conclusions: The results of our study showed that the G296S variant of the GATA4 gene was not associated with the development of CHD in Malaysian subjects. The use of HRM analysis proved a cost-effective, high-throughput, specific and sensitive genotyping technique which eliminates the need for unnecessary sequencing.

scholarly journals Identification of disease resistance-linked alleles in Vitis vinifera germplasm

2019 ◽  
Vol 13 ◽  
pp. 01004
Author(s):  
Maria Lucia Prazzoli ◽  
Silvia Lorenzi ◽  
Michele Perazzolli ◽  
Silvia Toffolatti ◽  
Osvaldo Failla ◽  
...  
Keyword(s):  
Vitis Vinifera ◽  
Ssr Markers ◽  
Genetic Resistance ◽  
Wine Quality ◽  
Breeding Programs ◽  
Relationship Analysis ◽  
Central Asian ◽  
Grape Varieties ◽  
Group Pattern ◽  
Grape Breeding

Introgression of genetic resistance to fungal diseases from American and Asian Vitis species traditionally pursued in grape breeding programs, although facilitated by molecular tools, has an impact on wine quality that still slows down development of competitive varieties. A contribution to the genetic improvement of grapevines for resistance to pathogens may come from unexplored genetic resources of the Eurasian Vitis vinifera L. In the present study, a hundred grapevine accessions from Georgia, Armenia and Azerbaijan were genotyped with SSR markers linked to QTLs for resistance to downy and powdery mildew, and with 21 SSR markers widely used for genetic diversity and relationship analysis. Looking at population genetic structure, Armenian and Azerbaijani accessions fell within the same cluster and were included among the Central Asian grape varieties of a homogeneous dataset, while Georgian accessions formed a separate group. Pattern of SSR alleles flanking the locus Ren1 and associated with resistance to Erysiphe necator in 'Kishmish vatkana', 'Dzhandzhal kara' and other Central Asian cultivars were found in three varieties from the Azerbaijani population that reached very high scores when assessed for PM resistance.

High-resolution melting (HRM) analysis as a feasible method for detecting spinal muscular atrophy via dried blood spots

2012 ◽  
Vol 413 (21-22) ◽  
pp. 1781-1785 ◽  
Author(s):  
Tze-Kiong Er ◽  
Tzu-Min Kan ◽  
Yu-Fa Su ◽  
Ta-Chih Liu ◽  
Jan-Gowth Chang ◽  
...  
Keyword(s):  
High Resolution ◽  
Spinal Muscular Atrophy ◽  
High Resolution Melting ◽  
Muscular Atrophy ◽  
Dried Blood Spots ◽  
Feasible Method ◽  
Blood Spots ◽  
Hrm Analysis
Sign in / Sign up

Export Citation Format

Share Document