Transcript quantification techniques usually rely on purified mRNAs. We report here a solution-based sandwich hybridization assay for the quantification of mRNAs from Escherichia coli without the need of prior RNA isolation. This assay makes use of four DNA oligonucleotide probes adjacently hybridizing to target RNA in clarified cell extracts. Two helper probes facilitate the hybridization of a...

leptin is secreted mainly by fat, which is involved in energy metabolism and reproduction. leptin and itsreceptor (ob-r) have been detected in human spermatozoa and testis, thus it can be concluded leptin involves in male physiology. the goal of this study was determination of the presence of leptin receptor mrna in the ram testis, epididymis and spermatozoa by reverse transcription polymerase ...

introduction: nicotinamide adenine dinucleotide phosphate (nadph) oxidase-2 (nox2) is one of the predominant sources of ros production during myocardial ischemia-reperfusion and can be induced by angiotensin ii. the evidence suggests that pharmacological blockers of renin-angiotensin system can exert direct tissue effects independent of their ability to regulate blood pressure. the mechanism(s)...

We describe a new strategy for quantifying mRNA by using polymerase chain reaction (PCR) coupled with HPLC. PCR-amplified products are directly analyzed with a specific HPLC column and are quantified by standardization against a housekeeping gene, beta-actin. To evaluate the experimental conditions, we examined the multidrug-resistance-associated mdr1 gene expression in two drug-sensitive cell ...

The physical manifestation of learning and memory formation in the brain can be expressed by strengthening or weakening of synaptic connections through morphological changes. Local actin remodeling underlies some forms of plasticity and may be facilitated by local β-actin synthesis, but dynamic information is lacking. In this work, we use single-molecule in situ hybridization to demonstrate tha...

MicroRNAs (miRNAs) are small, noncoding RNAs that negatively regulate gene expression. As miRNAs are involved in a wide range of biological processes and diseases, much effort has been invested in identifying their mRNA targets. Here, we present a novel combinatorial approach, RIP-chip-SRM (RNA-binding protein immunopurification + microarray + targeted protein quantification via selected reacti...

We developed a rapid, sensitive, and reproducible assay to quantify Candida albicans ACT1, CDR1, CDR2, ERG11, and MDR1 mRNA using a two-step reverse transcription and LightCycler real-time PCR (RT-LightCycler PCR) method with sequence-specific hybridization probes. We compared RT-LightCycler PCR with Northern hybridization for quantitative analysis of gene expression in isolates with various fl...

BACKGROUND Long-QT syndrome type 2 (LQT2) is caused by mutations in the human ether-a-go-go-related gene (hERG). More than 30% of the LQT2 mutations result in premature termination codons. Degradation of premature termination codon-containing mRNA transcripts by nonsense-mediated mRNA decay is increasingly recognized as a mechanism for reducing mRNA levels in a variety of human diseases. Howeve...

Pesticide effects on microbial community structure and activity in soil are reviewed, showing that methodological developments within the past few years have generated new possibilities for assessing pesticide effects. The first example is the use of mRNA quantification showing that nitrification processes are indeed very susceptible to some pesticides, and that there is correlation between the...

Delivery of neurotrophin-3 (NT-3) to severed sciatic nerves results in specific normalization of atrophied fast 2b gastrocnemius muscle fibres, and promotes preferential neuromuscular junction maturation of fast extensor digitorum longus (EDL). To investigate the selective influence on fast muscle reinnervation due to NT-3 delivery, we analyzed tyrosine kinase C (trkC) mRNA differential express...