Recently, more and more effort has been put into the miniaturization of genetic tests such as quantitative PCR (qPCR), because it is no doubt a powerful tool for molecular diagnosis and quantitative biology. In this paper, we developed a low density nanolitre droplet array generated on a chemical modified silicon chip for gene quantification. Reliable and sensitive two step real time qRT-PCR as...

As a gold standard for quantification of starting amounts of nucleic acids, real-time PCR is increasingly used in quantitative analysis of mtDNA copy number in medical research. Using supercoiled plasmid DNA and mtDNA modified both in vitro and in cancer cells, we demonstrated that conformational changes in supercoiled DNA have profound influence on real-time PCR quantification. We showed that ...

Polymerase chain reaction (PCR) is largely used in molecular biology for increasing the copy number of a specific DNA fragment. The succession of 20 replication cycles makes it possible to multiply the quantity of the fragment of interest by a factor of 1 million. The PCR technique has revolutionized genomics research. Several quantification methodologies are available to determine the DNA repl...

Next Generation Sequencing (NGS) is a powerful tool that depends on loading a precise amount of DNA onto a flowcell. NGS strategies have expanded our ability to investigate genomic phenomena by referencing mutations in cancer and diseases through large-scale genotyping, developing methods to map rare chromatin interactions (4C; 5C and Hi-C) and identifying chromatin features associated with reg...

.........................................................................................7 1. REVIEW OF THE LITERATURE.............................................................9 1.1. Prostate cancer...................................................................................9 1.2. Prostate specific antigen (PSA) and human glandular kallikrein (hK2)................11 1.2.1. Genes, mRNAs...

in present study, a sybr green based real time pcr assay was developed for specific detection and quantification of bacillus subtilis in dough used for bread making. new primer pairs were designed to amplify a 212 base pair fragment of the apre gene. specificity of these primer pairs was confirmed with conventional and real time pcr methods. standard curves constructed using the threshold cycle...

Digital polymerase chain reaction (PCR) is becoming ever more recognized amid the overwhelming revolution in DNA quantification, genomics, genetics, and diagnostics led by technologies such as next generation sequencing and studies at the single-cell level. The demand to quantify the amount of DNA and RNA has been driven to the molecular level and digital PCR, with its unprecedented quantificat...

Real-time PCR in nuclear ribosomal DNA (nrDNA) is becoming a well-established tool for the quantification of arbuscular mycorrhizal (AM) fungi, but this genomic region does not allow the specific amplification of closely related genotypes. The large subunit of mitochondrial DNA (mtDNA) has a higher-resolution power, but mtDNA-based quantification has not been previously explored in AM fungi. We...

Quantitative real-time or kinetic RT-PCR is increasingly used for the quantification of specific mRNA targets, especially in clinical applications. To quantify the mRNA of cytokines and their receptors, which play important roles in the pathogenesis of autoimmune diseases such as multiple sclerosis, we have developed quantitative two-step RT-PCR assays for IL-4, IL-4R, IFN-gamma, IFN-beta, and ...

BACKGROUND DzyNA-PCR is a general strategy for the detection and quantification of specific genetic sequences associated with disease or the presence of foreign agents. The method allows homogeneous gene amplification coupled with signal detection in a single closed vessel. METHODS The strategy involves in vitro amplification of genetic sequences using a DzyNA primer that harbors the compleme...