Fluorescent nucleic acid detection in polymerase chain reaction (PCR) generally uses oligonucleotide probes labeled with covalently attached dyes. However, unlabeled oligonucleotides in the presence of saturating DNA dyes can also serve as hybridization probes. The DNA dye, LCGreen Plus, and a 3'-blocked unlabeled probe are added before amplification, and asymmetric PCR is performed at a 1:5 to...

A strategy combining dual-suppression PCR and thermal asymmetric interlaced PCR was used to determine sequences flanking microsatellite regions in Pythium helicoides. The primer pairs were designed to amplify loci containing (AC)n, (GA)n, (AGC)n, (CAC)n(CAA)n, (TCA)n and (CTTT)n repeats from the P. helicoides nuclear genome. The PCR products of each primer pair, amplified from three representat...

1.Barik, S. and M.S. Galinski. 1991. “Megaprimer” method of PCR: increased template concentration improves yield. BioTechniques 10:489-490. 2.Colosimo, A., Z. Xu, G. Novelli, B. Dallapiccola, and D.C. Gruenert. 1999. Simple version of “megaprimer” PCR for site-directed mutagenesis. BioTechniques 26:870-873. 3.Datta, A.K. 1995. Efficient amplification using “megaprimer” by asymmetric polymerase ...

Microarrays provide a powerful analytical tool for the simultaneous detection of multiple pathogens. We developed diagnostic suspension microarrays for sensitive and specific detection of the biothreat pathogens Bacillus anthracis, Yersinia pestis, Francisella tularensis and Coxiella burnetii. Two assay chemistries for amplification and labeling were developed, one method using direct hybridiza...

We describe a method to screen pools of DNA from multiple transposon lines for insertions in many genes simultaneously. We use thermal asymmetric interlaced-PCR, a hemispecific PCR amplification protocol that combines nested, insertion-specific primers with degenerate primers, to amplify DNA flanking the transposons. In reconstruction experiments with previously characterized Arabidopsis lines ...

It has been shown that fluorescence resonance energy transfer (FRET)-based PCR, including the TaqMan assay and molecular beacons, has potential for rapid detection of pathogens. In these promising real-time detection assays a single internal oligonucleotide probe labeled on both the 5' (reporter) and 3' (quencher) ends is used for selective generation of fluorescence. In this paper, we describe...

DNA nanoengineering, in particular, origami has potential applications a variety of areas including, for example, nanoelectronics, biomedical diagnostics, and therapeutics. To fully realize the self-assembly these other areas, methods must be available economical, scalable, reliable production single-stranded (ssDNA) scaffolds from virtually any source. In this review, we will describe virtues ...

conclusions our underlying hypothesis is that recapitulating the three-dimensional early embryonic structure, in order to reproduce better in vitro the three-dimensional morphogenetic-like re-arrangements, would improve cells differentiation, when in vivo transplanted; moreover, it could be used as a simplified cancer disease model and reliable drug evaluation method as well. results the proces...