Structural Key Bit Occurrence Frequencies and Dependencies in PubChem and Their Effect on Similarity Searches.

Little published literature exists on the 881 bit structural keys used by PubChem for categorizing and comparing the compounds present in its database. We characterized these structural keys by examining their frequencies of occurrence within the PubChem compound database. In addition, bit dependencies, defined as the universal presence of a bit given the presence of another, were determined.

Rapid and direct microRNA quantification by an enzymatic luminescence assay.

A quantitative bioluminescence assay for rapid and sensitive microRNA (miRNA) expression analysis was developed. The assay uses miRNA directly as a primer for binding to a circular single-stranded DNA template, followed by rolling circle amplification. The detection of inorganic pyrophosphate (PPi) molecules released during the DNA polymerization and amplification process is performed by a multi-enzyme system.

Transient model of thermal deactivation of enzymes.

The kinetics of enzyme deactivation provide useful insights on processes that determine the level of biological function of any enzyme. Photinus pyralis (firefly) luciferase is a convenient enzyme system for studying mechanisms and kinetics of enzyme deactivation, refolding, and denaturation caused by various external factors, physical or chemical by nature. In this report we present a study of luciferase deactivation caused by increased temperature (i.

Efficiency and specificity of microRNA-primed nucleotide analog incorporation by various DNA polymerases.

MicroRNAs (miRNAs) as endogenous regulators of gene expression have spurred a surge of interest for their quantification and expression analysis. High-sensitivity and high-specificity miRNA detection techniques, such as real-time polymerase chain reaction and recently introduced bioluminescent miRNA detection, require systematic study of DNA polymerases for use with miRNAs. In this study, a variety of DNA polymerases were studied to assess their capabilities of using miRNA as a primer and incorporating 2'-deoxyadenosine-5'-O-(1-thiotriphosphate) as a dATP alternative during DNA strand extension.

Rapid and quantitative quality control of microarrays using cationic nanoparticles.

The fabrication quality of microarrays significantly influences the accuracy and reproducibility of microarray experiments. In this report, we present a simple and fast quality control (QC) method for spotted oligonucleotide and cDNA microarrays. It employs a nonspecific electrostatic interaction of colloidal gold nanoparticles with the chemical groups of DNA molecules and other biomolecules immobilized on the microarray surface that bear positive or negative charges.

Hybridization kinetics on microarray surfaces.

Kinetic hybridization data are compared to a number of different models. A first-order Langmuir model provides the best fit to the data.

A multienzyme bioluminescent time-resolved pyrophosphate assay.

We have developed a high-sensitivity assay for measurement of inorganic pyrophosphate (PPi) in adenosine 5'-triphosphate (ATP)-contaminated samples. The assay is based on time-resolved measurements of the luminescence kinetics and implements multiple enzymes to convert PPi to ATP that is, in turn, utilized to produce light and to hydrolyze PPi for measurement of the steady state background luminescence. A theoretical model for describing luminescence kinetics and optimizing composition of the assay detection mixture is presented.

Label-free detection of biomolecules on microarrays using surface-colloid interaction.

Binding of charged nanoparticles to nonmodified DNA and RNA target molecules on microarrays allows detection of these nucleic acids by locating bound nanoparticles on microarray surface. Two computational models have been developed to study the ionic interaction of colloidal particles and biopolymer molecules on a microarray surface. One model represents a basic approach based on mass action law for simulation charge effects versus the chemical composition of the interacting array surface and nanoparticle.

Microarray gene expression analysis free of reverse transcription and dye labeling.

A new microarray system has been developed for gene expression analysis using cationic gold nanoparticles with diameters of 250 nm as a target detection reagent. The approach utilizes nonlabeled target molecules hybridizing with complementary probes on the array, followed by incubation in a colloidal gold solution. The hybridization signal results from the precipitation of nanogold particles on the hybridized spots due to the electrostatic attraction of the cationic gold particles and the anionic phosphate groups in the target DNA backbone.