Instrumentation, chemistry, and software for high-throughput genotyping using fluorescent melting curves are described. The LightTyper system provides post-amplification genotyping within 10 min using samples in 96- or 384-well microplate formats. The system is homogenous because all reagents are added at the beginning of the reaction and there is no sample manipulation between amplification an...

Real-time monitoring of PCR is one of the most important methods for DNA and RNA detection widely used in research and medical diagnostics. Here we describe a new approach for combined real-time PCR monitoring and melting curve analysis using a 3' end-blocked Exciton-Controlled Hybridization-sensitive fluorescent Oligonucleotide (ECHO) called Eprobe. Eprobes contain two dye moieties attached to...

OBJECTIVE The goal of this research was to develop a screening instrument for early identification among hospitalized children of medical child abuse (MCA). METHODS We developed a preliminary screening instrument for the early identification of MCA. Items were chosen based on published characteristics of MCA, including caregiver, patient, and illness information. Each item in the instrument w...

25 A single-tube method – ligation mediated real-time PCR high-resolution melt analysis 26 (LMqPCR HRMA) – was modified for rapid typing of ESKAPE-pathogens. A 97 % 27 agreement (60/62 isolates) was achieved in comparison to PFGE which indicates that 28 LMqPCR HRMA is a rapid and accurate screening tool to monitor for nosocomial outbreaks. 29 on O cber 1, 2017 by gest ht://jcm .sm .rg/ D ow nla...

A rapid competitive PCR method was developed to quantify DNA on the LightCycler. It rests on the quantitative information contained in the melting curves obtained after amplification in the presence of SYBR Green I. Specific hybridization probes are not required. Heterologous internal standards sharing the same primer binding sites and having different melting temperatures to the natural PCR pr...

BACKGROUND Single-base pair missense mutations in exons 10, 11, 13, 14, 15, and 16 of the RET protooncogene are associated with the autosomal dominant multiple endocrine neoplasia type 2 (MEN2) syndromes: MEN2A, MEN2B, and familial medullary thyroid carcinoma. The current widely used approach for RET mutation detection is sequencing of the exons. METHODS Because RET mutations are rare and the...

Melting fine structure of the nuclear DNA isolated from the filamentous fungus Fusarium graminearum Schwabe is presented. Optical melting profiles of nuclear DNA were analyzed by using a combination of curve fitting and derivative techniques. The "melting components" were obtained from the derivative curve by a simple decomposition technique. Differential optical melting curves of unsheared nuc...