Transl Androl Urol, Vol 3, Suppl 1 September 2014 www.amepc.org/tau © Translational Andrology and Urology. All rights reserved. Objective: To establish a high resolution melting analysis with unlabeled probes for genotyping CCL2-2518 T > C. Methods: Two unlabeled probes were designed. One is complementary to wild type and another is complementary to the mutant type. A total of 71 health people ...

Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) is the method of choice for rapid and reproducible measurements of cytokine or growth factor expression in small samples. Fluorescence detection methods for monitoring real-time PCR include fluorogenic probes labelled with reporter and quencher dyes, such as Taqman probes or Molecular Beacons and the dsDNA-binding d...

The data in this article are related to the research article entitled "Optimization of melting analysis with TaqMan probes for detection of KRAS, NRAS, and BRAF mutations" Botezatu et al. [1]. Somatic mutations in the PIK3CA gene ("hot spots" in exons 9 and 20) are found in many human cancers, and their presence can determine prognosis and a treatment strategy. An effective method of mutation s...

Supplementary material 1 Real-time quantitative PCR: Sequences of the primers and TaqMan probe for ADAMTS1, 4, 5, 8, 9 and 15 were designed using Primer Express software (Applied Biosystems, Foster City, CA): forward primer 5’-CCATCCCAAGAGTATCACATGTCT-3’, reverse primer 5’-CACTATGACACAGCAATTCTTTTCAC-3’ and TaqMan probe 5’-FAM-CCCACACAAGTCCTGTC-MGB-3 for ADAMTS1; forward primer 5’-TCACTGACTTCCTG...

In this review, we report on patents concerning self-quenching DNA probes for assaying DNA during or after amplification as well as for direct assaying DNA or RNA, for example in living cells. Usually the probes consist of fluorescently labeled oligonucleotides whose fluorescence is quenched in the absence of the matching target DNA. Thereby the fluorescence quenching is based on fluorescence r...

An important consideration in the design of oligonucleotide probes for homogeneous hybridization assays is the efficiency of energy transfer between the fluorophore and quencher used to label the probes. We have determined the efficiency of energy transfer for a large number of combinations of commonly used fluorophores and quenchers. We have also measured the quenching effect of nucleotides on...

TaqMan genotyping with real-time PCR is a reliable method for single nucleotide polymorphism detection, which is done by probes. These oligonucleotides should be short enough to avoid mismatch hybridization, as well as having 5-10°C higher melting temperature than the primers of real-time PCR reaction. One approach for these qualities is to conjugate the probe with minor groove binder (MGB). Ha...

A quantitative real-time TaqMan PCR assay for detection of human adenoviruses (HAdV) was developed using broadly reactive consensus primers and a TaqMan probe targeting a conserved region of the hexon gene. The TaqMan assay correctly identified 56 representative adenovirus prototype strains and field isolates from all six adenovirus species (A to F). Based on infectious units, the TaqMan assay ...