Storage conditions are considered to be a critical component of DNA-based microbial community analysis methods. However, whether differences in short-term sample storage conditions impact the assessment of bacterial community composition and diversity requires systematic and quantitative assessment. Therefore, we used barcoded pyrosequencing of bacterial 16S rRNA genes to survey communities, ha...

This article describes a dispensation order generation algorithm for genotyping using the Pyrosequencing method. The input template of the algorithm is a slightly restricted regular expression over the DNA strings that can be expected in a given sample. The algorithm computes a dispensation order that allows for determining, for each polymorphism in the input template, the genotype of any sampl...

BACKGROUND The Human Microbiome Project has ushered in a new era for human metagenomics and high-throughput next-generation sequencing strategies. CONTENT This review describes evolving strategies in metagenomics, with a special emphasis on the core technology of DNA pyrosequencing. The challenges of microbial identification in the context of microbial populations are discussed. The developme...

Manipulating the mouse genome is a widespread technology with important applications in many biological fields ranging from cancer research to developmental biology. Likewise, correlations between copy number variations (CNVs) and human diseases are emerging. We have developed the reference-query pyrosequencing (RQPS) method, which is based on quantitative pyrosequencing and uniquely designed p...

Aims: Enhancer of Zeste Homolog 2 (EZH2) is a critical enzymatic subunit of Polycomb Repressive Complex 2 (PRC2), which provides inhibitory regulation of gene transcription through trimethylation of histone H3 on lysine 27 (H3K27me3). The somatic mutation EZH2 Y641 in its highly conserved catalytic SET domain has been reported in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL...

The results of investigations into performing DNA sequencing chemistry on a picoliter-scale electrowetting digital microfluidic platform are reported. Pyrosequencing utilizes pyrophosphate produced during nucleotide base addition to initiate a process ending with detection through a chemiluminescence reaction using firefly luciferase. The intensity of light produced during the reaction can be q...

BACKGROUND Pyrosequencing is a DNA sequencing technology based on the sequencing-by-synthesis principle. METHODS The technique is built on a 4-enzyme real-time monitoring of DNA synthesis by bioluminescence using a cascade that upon nucleotide incorporation ends in a detectable light signal (bioluminescence). The detection system is based on the pyrophosphate released when a nucleotide is int...