Removal of artifact bias from qPCR results using DNA melting curve analysis

High-resolution melting-curve analysis of ligation-mediated qPCR;

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...

Rapid competitive PCR using melting curve analysis for DNA quantification.

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...

In-tube DNA methylation profiling by fluorescence melting curve analysis.

BACKGROUND Most PCR assays for detection of 5-methylcytosine in genomic DNA entail a two-step procedure, comprising initial PCR amplification and subsequent product analysis in separate operations that usually require manual transfer. These methods generally provide information about methylation of only a few CpG dinucleotides within the target sequence. METHODS An in-tube methylation assay i...

Comparison of cattle BoLA-DRB3 typing by PCR-RFLP, direct sequencing, and high-resolution DNA melting curve analysis

Major histocompatibility complex (MHC) represents an important genetic marker for manipulation to improve the health and productivity of cattle. It is closely associated with numerous disease susceptibilities and immune responses. Bovine MHC, also called bovine leukocyte antigen (BoLA), is considered as a suitable marker for genetic diversity studies. In cattle, most of the polymorphisms are lo...

Reversal of PCR bias for improved sensitivity of the DNA methylation melting curve assay.