Novel methods were developed to isothermally detect target nucleic acids and initiate signal amplification cascades. The methods utilize target-specific MNAzymes to activate universal primer molecules. These primers can subsequently promote the autonomous synthesis of DNAzymes capable of cleaving nucleic acid substrates and generating signal, which further creates the potential for circular fee...

We have developed a new method for high-throughput genotyping of single nucleotide polymorphisms (SNPs). The technique involves PCR amplification of genomic DNA with two tailed allele-specific primers that introduce priming sites for universal energy-transfer-labeled primers. The output of red and green light is conveniently scored using a fluorescence plate reader. The new method, which was va...

The use and validation of a strategy that allows a universal set of bar-coded sequencing primers to be appended to an amplified PCR product is described. The strategy allows a modular approach, in that the same bar code can be used with two or more target-specific primer sets, even simultaneously.

DNA primer sets, labeled with two fluorescent dyes to exploit fluorescence energy transfer (ET), can be efficiently excited with a single laser line and emit strong fluorescence at distinctive wavelengths. Such ET primers are superior to single fluorophore-labeled primers for DNA sequencing and other multiple color-based analyses [J. Ju, C. Ruan, C. W. Fuller, A. N. Glazer and R. A. Mathies (19...

Biases on molecular techniques used to study microbial communities in the anammox process are not always considered. This work aimed evaluate different approaches bacterial an reactor treating real urban wastewater at mainstream conditions. 16S rRNA amplicon sequencing with primer sets, metagenomics, and qPCR were evaluated. An enriched feed synthetic was as a positive control. We reported that...

The diversity of protozoan-associated methanogens in cattle was investigated using five universal archaeal small-subunit (SSU) rRNA gene primer sets. Methanobrevibacter spp. and rumen cluster C (distantly related to Thermoplasma spp.) were predominant. Significant differences in species composition among libraries indicate that some primers used previously to characterize rumen methanogens exhi...

We investigated the bacterial communities of nine Bartonella-positive fleas (n = 6 Oropsylla hirsuta fleas and n = 3 Oropsylla montana fleas), using universal primers, clone libraries, and DNA sequencing. DNA sequences were used to classify bacteria detected in a phylogenetic context, to explore community assembly patterns within individual fleas, and to survey diversity patterns in dominant li...

development of yeast artificial chromosome (y ac) vectors, molecular cloning of large segments of chromosomal dna, and their propagation in yeast cells has become feasible. overlapping y ac provides a route to the development of physical maps of entire mammalian chromosomes. a rapid method was developed to isolate and sequence termini of y ac inserts quickly. the y ac clone is digested with a r...

A 40-year-old healthy male employed in a bakery presented with a single lung nodule and underwent investigations to rule out pulmonary carcinoma. Biopsy was positive for yeast cells, which did not match common fungal pathogens. PCR assay of paraffin-embedded tissue and nucleotide sequencing with ribosomal ITS1-ITS2 universal primers revealed the presence of Saccharomyces cerevisiae.