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high resolution melt

Quanyou Feng, Lei Yang, Yongliang Zhong, Dong Guo, Guoliang Liu, Linghai Xie, Wei Huang, Rong Tong
Biodegradable polyesters with various tacticities have been synthesized by means of stereoselective ring-opening polymerization of racemic lactide and β-lactones but with limited side-chain groups. However, stereoselective synthesis of functional polyesters remains challenging from O-carboxyanhydrides that have abundant pendant side-chain functional groups. Herein we report a powerful strategy to synthesize stereoblock polyesters by stereoselective ring-opening polymerization of racemic O-carboxyanhydrides with the use of photoredox Ni/Ir catalysts and a selected Zn complex with an achiral ligand...
April 19, 2018: Nature Communications
Marjanu Hikmah Elias, Husin Azlan, Abdul Aziz Baba, Ravindran Ankathil
BACKGROUND: In exploring the cause of Imatinib mesylate (IM) resistance among chronic myeloid leukemia (CML) patients who do not harbouring BCR-ABL dependent mechanism, BCR-ABL independent pathways are the most possible pathways that should be explored. In BCR-ABL independent pathway, SOCS1 plays an important role as it helps to regulate optimal JAK/STAT activity. OBJECTIVE: To identify the association of SOCS1 gene hypermethylation in mediating IM Resistance. METHOD: The SOCS1 promoter methylation level of 92 BCR-ABL non mutated IM resistant CML patients, 83 IM good response CML patients and 5 normal samples from healthy individuals were measured using Methylation Specific- High Resolution Melt (MS-HRM) analysis...
April 18, 2018: Cardiovascular & Hematological Disorders Drug Targets
Yue Fu, Wen-Jun Liu
Thalassemia laboratory diagnostic techniques include screening test and genetic test, the former diagnosis method is mainly based on the morphology of red blood cells and the physical and chemical properties, including routine analysis of blood, red blood cell morphology test, red blood cell permeability test, hemoglobin component analysis, etc. The latter is mainly based on various of molecular biology experiments developed on the basis of PCR, including gap-PCR,PCR-RDB,melting curve analysis,high resolution melting analysis,real-time PCR,gene chip and DNA sequencing, etc...
April 2018: Zhongguo Shi Yan Xue Ye Xue za Zhi
Lu-Lu Sun, Yuan Liu, Xiao Sun, Lei Pan, Dan Wu, Yu-Dong Wang
Background: Promoter methylation of MGMT and C13ORF18 has been confirmed as a potential biomarker for early diagnosis of cervical cancer. The aim of this study was to evaluate the performance of MGMT and C13ORF18 promoter methylation for triage of cytology screening samples and explore the potential mechanism. Methods: Methylation-sensitive high-resolution melting was used to detect promoter methylation of MGMT and C13ORF18 in 124 cervical samples. High-risk human papillomavirus (HR-HPV) was detected by the Digene Hybrid Capture 2® ...
April 20, 2018: Chinese Medical Journal
Yan Zhang, Haibo Zhou, Hongxu Sun, Jie Chen, Di Huang, Xu Han, Xiyun Ren, Shangqun Lin, Qing Fan, Wenjing Tian, Yashuang Zhao
KIBRA was reported to be involved in various types of cancer and can be detected in blood. The purpose of this study was to investigate the relationship between the status of KIBRA methylation in peripheral blood leukocytes and gastric cancer (GC) risk. A case-control study was carried out to evaluate the association of blood cell-derived KIBRA methylation with the risk of GC using methylation-sensitive high-resolution melting analysis. A total of 393 cases and 393 controls were detected, respectively. Compared with the subjects in the KIBRA negative methylation (NM) group, positive methylation (PM) subjects exhibited a 1...
April 16, 2018: Cancer Medicine
Wanying Zhang, Tao Wang, Shuaiwu Huang, Xiuli Zhao
OBJECTIVE: To detect mutation of HPGD gene among three pedigrees affected with primary hypertrophic osteoarthropathy (PHO) by DNA sequencing and high-resolution melting (HRM) analysis. METHODS: Genomic DNA was extracted from peripheral blood samples collected from the pedigrees. PCR and direct sequencing were carried out to identify potential mutations of the HPGD gene. Amplicons containing the mutation spot were generated by nested PCR. The products were then subjected to HRM analysis using the HR-1 instrument...
April 10, 2018: Zhonghua Yi Xue Yi Chuan Xue za Zhi, Zhonghua Yixue Yichuanxue Zazhi, Chinese Journal of Medical Genetics
Mridu Sinha, Hannah Mack, Todd P Coleman, Stephanie I Fraley
DNA melting analysis provides a rapid method for genotyping a target amplicon directly after PCR amplification. To transform melt genotyping into a broad-based profiling approach for heterogeneous samples, we previously proposed the integration of universal PCR and melt analysis with digital PCR. Here, we advanced this concept by developing a high-resolution digital melt platform with precise thermal control to accomplish reliable, high-throughput heat ramping of microfluidic chip digital PCR reactions. Using synthetic DNA oligos with defined melting temperatures, we characterized sources of melting variability and minimized run-to-run variations...
April 1, 2018: SLAS Technology
Sónia Gomes, Cláudia Castro, Sara Barrias, Leonor Pereira, Pedro Jorge, José R Fernandes, Paula Martins-Lopes
The wine sector requires quick and reliable methods for Vitis vinifera L. varietal identification. The number of V. vinifera varieties is estimated in about 5,000 worldwide. Single Nucleotide Polymorphisms (SNPs) represent the most basic and abundant form of genetic sequence variation, being adequate for varietal discrimination. The aim of this work was to develop DNA-based assays suitable to detect SNP variation in V. vinifera, allowing varietal discrimination. Genotyping by sequencing allowed the detection of eleven SNPs on two genes of the anthocyanin pathway, the flavanone 3-hydroxylase (F3H, EC: 1...
April 11, 2018: Scientific Reports
Sihai Luo, Liucheng Zhou, Xuede Wang, Xin Cao, Xiangfan Nie, Weifeng He
As an innovative surface technology for ultrahigh strain-rate plastic deformation, laser shock peening (LSP) was applied to the dual-phase TC11 titanium alloy to fabricate an amorphous and nanocrystalline surface layer at room temperature. X-ray diffraction, transmission electron microscopy, and high-resolution transmission electron microscopy (HRTEM) were used to investigate the microstructural evolution, and the deformation mechanism was discussed. The results showed that a surface nanostructured surface layer was synthesized after LSP treatment with adequate laser parameters...
April 6, 2018: Materials
Karen M Jiménez, Diego A Forero
Alterations in telomere length (TL) have been associated with several diseases and a method based on qPCR, the Monochrome Multiplex Real-Time Quantitative PCR (MMQPCR) technique, has been used extensively for the analysis of TL. Some previous studies have been found that certain methodological conditions can affect the measurement of TL. The aim of the study was to evaluate the performance of eight different commercially available SYBR Green and High-Resolution Melting (HRM) mixes on the measurement of TL by the MMQPCR method...
April 5, 2018: Molecular Biology Reports
Paulo Roberto Xavier Tomaz, Juliana Rocha Santos, Jaqueline Scholz, Tânia Ogawa Abe, Patrícia Viviane Gaya, André Brooking Negrão, José Eduardo Krieger, Alexandre Costa Pereira, Paulo Caleb Júnior Lima Santos
BACKGROUND: The identification of variants in the nicotinic acetylcholine receptor (nAChR) subunit genes associated with smoking phenotypes are increasingly important for prevention and treatment of nicotine dependence. In the context of personalized medicine, the aims of this study were to evaluate whether cholinergic receptor nicotinic alpha 2 (CHRNA2), cholinergic receptor nicotinic alpha 3 (CHRNA3), cholinergic receptor nicotinic alpha 5 (CHRNA5) and cholinergic receptor nicotinic beta 3 (CHRNB3) polymorphisms were associated with nicotine dependence severity, and to investigate possible pharmacogenetics markers of smoking cessation treatment...
April 5, 2018: BMC Medical Genetics
Banya Banowary, Van Tuan Dang, Subir Sarker, Joanne H Connolly, Jeremy Chenu, Peter Groves, Shane Raidal, Seyed Ali Ghorashi
Campylobacter infection is a common cause of bacterial gastroenteritis in humans and remains a significant global public health issue. The capability of two multiplex PCR (mPCR)-high-resolution melt (HRM) curve analysis methods (i.e., mPCR1-HRM and mPCR2-HRM) to detect and differentiate 24 poultry isolates and three reference strains of Campylobacter jejuni and Campylobacter coli was investigated. Campylobacter jejuni and C. coli were successfully differentiated in both assays, but the differentiation power of mPCR2-HRM targeting the cadF gene was found superior to that of mPCR1-HRM targeting the gpsA gene or a hypothetical protein gene...
March 2018: Avian Diseases
Faranak Rezaei, Mehri Haeili, Abbasali Imani Fooladi, Mohammad Mehdi Feizabadi
Objectives: Development of molecular techniques for rapid detection of drug resistant tuberculosis allows for the prompt initiation of appropriate anti-TB treatment. We aimed to assess high-resolution melting (HRM) analysis for the detection of rpsL, rrs and embB mutations to identify streptomycin and ethambutol resistance in Mycobacterium tuberculosis. Material and Methods: A total of 76 clinical isolates of M. tuberculosis including 25 SM-R, 21 EB-R and 30 drug susceptible - determined by the proportion method of drug susceptibility testing (DST) - were analyzed by HRM analysis, and the results were confirmed using DNA sequencing...
December 2017: Mædica
Junying Sun, Gali Bingga, Zhicheng Liu, Chunhong Zhang, Haiyan Shen, Pengju Guo, Jianfeng Zhang
Differentiation of classical strains and highly pathogenic strains of porcine reproductive and respiratory syndrome virus is crucial for effective vaccination programs and epidemiological studies. We used nested PCR and high resolution melting curve analysis with unlabeled probe to distinguish between the classical and the highly pathogenic strains of this virus. Two sets of primers and a 20 bp unlabeled probe were designed from the NSP3 gene. The unlabeled probe included two mutations specific for the classical and highly pathogenic strains of the virus...
March 30, 2018: Molecular and Cellular Probes
Angel Aleman, Chao Li, Hicham Zaid, Hanna Kindlund, Joshua Fankhauser, Sergey V Prikhodko, Mark S Goorsky, Suneel Kodambaka
Pd(111) thin films, ∼245 nm thick, are deposited on Al2 O3 (0001) substrates at ≈0.5 T m , where T m is the Pd melting point, by ultrahigh vacuum dc magnetron sputtering of Pd target in pure Ar discharges. Auger electron spectra and low-energy electron diffraction patterns acquired in situ from the as-deposited samples reveal that the surfaces are compositionally pure 111-oriented Pd. Double-axis x-ray diffraction (XRD) ω-2θ scans show only the set of Pd 111 peaks from the film. In triple-axis high-resolution XRD, the full width at half maximum intensity Γω of the Pd 111 ω-rocking curve is 630 arc sec...
May 2018: Journal of Vacuum Science & Technology. A, Vacuum, Surfaces, and Films
Dong-Mei Fu, Yu-Lin Zhou, Jing Zhao, Ping Hu, Zheng-Feng Xu, Shi-Ming Lv, Jun-Jie Hu, Zhong-Min Xia, Qi-Wei Guo
Klinefelter syndrome (KS) is the set of symptoms that result from the presence of an extra X chromosome in males. Postnatal population-based KS screening will enable timely diagnosis of this common chromosomal disease, providing the opportunity for early intervention and therapy at the time point when they are most effective and may prevent later symptoms or complications. Therefore, through this study, we introduced a simple high-resolution melting (HRM) assay for KS screening and evaluated its clinical sensitivity and specificity in three medical centers using 1373 clinical blood samples...
March 30, 2018: Asian Journal of Andrology
Farhadul Islam, Johnny C Tang, Vinod Gopalan, Alfred K Lam
The aberrant DNA methylation has been noted to occur at promoter of tumor suppressor, cell adhesion, DNA repair, and other growth regulating genes during the progression of nonneoplastic esophageal mucosa to Barrett esophagus to esophageal adenocarcinoma. Methylation-mediated silencing of individual gene or concurrent loss of a number of genes plays crucial roles in dysplasia-metaplasia-neoplasia sequence of esophageal adenocarcinoma. In addition, promoter methylation of genes had shown significant prognostic potential in patients with esophageal adenocarcinoma...
2018: Methods in Molecular Biology
Anders Abildgaard, Sara K Tovbjerg, Axel Giltay, Liselot Detemmerman, Peter H Nissen
BACKGROUND: The lactase persistence phenotype is controlled by a regulatory enhancer region upstream of the Lactase (LCT) gene. In northern Europe, specifically the -13910C > T variant has been associated with lactase persistence whereas other persistence variants, e.g. -13907C > G and -13,915 T > G, have been identified in Africa and the Middle East. The aim of the present study was to compare a previously developed high resolution melting assay (HRM) with a novel method based on loop-mediated isothermal amplification and melting curve analysis (LAMP-MC) with both whole blood and DNA as input material...
March 26, 2018: Clinica Chimica Acta; International Journal of Clinical Chemistry
Karen M Jiménez, Angela J Pereira-Morales, Diego A Forero
BACKGROUND: Epigenetic factors have been identified in the past years as interesting candidates for psychiatric disorders and related endophenotypes. It has been found that the methylenetetrahydrofolate reductase (MTHFR) gene is associated with major depressive disorder, and the aim of the current study was to examine the possible association between perceived stress and MTHFR methylation, taking into account depressive symptoms as a covariate. PARTICIPANTS AND METHODS: Seventy-eight healthy Colombian participants (mean age=20...
March 27, 2018: Psychiatric Genetics
Priyanka Mishra, Ashutosh K Shukla, Velusamy Sundaresan
Senna alexandrina (Fabaceae) is a globally recognized medicinal plant for its laxative properties as well as the only source of sennosides, and is highly exported bulk herb from India. Its major procurement is exclusively from limited cultivation, which leads to risks of deliberate or unintended adulteration. The market raw materials are in powdered or finished product form, which lead to difficulties in authentication. Here, DNA barcode tags based on chloroplast genes ( rbcL and matK ) and intergenic spacers ( psbA-trnH and ITS ) were developed for S...
2018: Frontiers in Plant Science
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