生物化学技术

There is a great deal of interest in analytical biochemistry in the separation and identification of biologically important polymers, such as DNA protein and complex carbohydrate molecules (1,2). For relatively short single-stranded DNA units (i.e., oligonucleotides) and carboh ...

Many papers have appeared concerning the application of capillary electrophoresis (CE) to the resolution of chiral compounds. Predominantly, separations have been achieved in Free Solution Capillary Electrophoresis (FSCE) employing cyclodextrins (CDs) as chiral select ...

In conventional liquid chromatography, the flow of eluent is generated by the application of a pressure gradient, which is usually provided by means of a high-pressure mechanical pump. However, under certain conditions, the mobile phase can be forced to migrate through the chromatograph ...

An important part of the ribozyme efficiency-screening process is to have a fast and accurate way to measure steady-state levels of the target RNA. Here, we describe the use of real-time polymerase chain reaction (PCR) for quantification of ribozyme target transcripts. In contrast to classi ...

Nitric oxide synthase (NOS) catalyzes the conversion of L-arginine, molecular oxygen, and nicotinamide adenine dinucleotide phosphate (NADPH) to NO, citrulline, and NADP+ (reviewed in ref. 1). This chapter describes the measurement of NOS activity by utilizing the conversion of oxyh ...

To date, the cDNAs and genes encoding three distinct forms of the enzyme nitric oxide synthase (NOS) have been described (reviewed in ref. 1). The family of NOS enzymes generate nitric oxide (NO) from arginine, producing citrulline as a co-product and requires the presence of heme, tetrahydrobio ...

We describe techniques for identifying functional promoter elements in the 5′-flanking region of the murine cyclic nucleotide phosphodiesterase 3B (mPDE3B) gene. The 5′-flanking region of the mPDE3B gene was cloned and sequenced, and putative transcription factor binding sites w ...

2′-O-Alkyloligoribonucleotides possess properties that render them ideally suited for studying RNA processing and for the affinity chromatography of RNA-protein complexes. Particularly useful compounds are those in which alkyl is methyl (1–7), ethyl (8), allyl (9), or butyl (10), wi ...

Monitoring conversion of radiolabeled arginine to citrulline, the co-product of the nitric oxide synthase (NOS)-catalyzed reaction, is a widely used method to assay for NOS activity. In cases where NOS is the sole or overwhelmingly predominant arginine-metabolizing enzyme present ...

The involvement of nitric oxide (NO) as an effector molecule in many physiological and pathological situations is now firmly established. Although NO can be measured directly using NO-specific electrodes and chemiluminescence, the measurement of the stable end-products of NO meta ...

Noncultural methods are available for the direct detection of various microorganisms in clinical specimens and/or for the identification of certain microbial agents recovered from culture. The Gen-Probe PACE 2 (Probe Assay—Chemiluminescence Enhanced) system for Neisseria ...

Nowadays, several chemicals are being assayed to control the proliferation of the ascomycete Ascosphera apis in honey beehives. The presence and growing of these fungi in bee larvae causes their death and as a result of this a reduction in the number of bees is appreciated in the colony, and this also pr ...

The presence in the body of drugs of abuse can be identified by a variety of laboratory procedures. The standard in drug testing is the immunoassay screen, followed by a gas chromatography/mass spectrometry (GC/MS) confirmation conducted on a urine sample. In general, drug concentrations in u ...

The purpose of this chapter is to describe a supercritical fluid extraction (SFE) method coupled with enzyme immunoassay analysis (EIA) for the determination of the herbicides: 2,4-dichlorophenoxyacetic acid (2,4-D), simazine, atrazine, alachlor, and trifluralin in soil.

The first report demonstrating the feasibility of supercritical fluids as solvent media for performing enzymatic reactions was published in 1986 (1). Since then several reports have confirmed that the relatively low critical temperature and pressure of supercritical fluid carb ...

The analysis of drugs and their metabolites in biological fluids represents an essential role in pharmaceutical and toxicology studies. High performance liquid chromatography (HPLC) has emerged as a particularly powerful analytical technique for drug analysis since many water ...

Beta-blockers have some clinical use in the treatment of angina pectoris, hypertension, and tachycardia. In addition, they have been used to control migraine, chronic alcoholism, schizophrenia, essential tremor, and cardiac effects of cocaine overdose. Unfortunately, misuse of t ...

The determination of trace organic compounds in aqueous samples usually involves isolation of the fraction of interest followed by subsequent separation by means of a chromatographic technique. When high resolution is needed, the main chromatographic techniques usually emplo ...

Malaria is a major disease in many countries and, according to an estimation by the World Health Organization (WHO), approx 300–500 million people contract malaria yearly and almost 2 million die annually (1). Controlling malaria is now becoming very problematic because of the developing re ...

Pulsed-field gel electrophoresis (PFGE) has been successfully used to generate physical maps of regions of human chromosomes near genes or single copy probes. However, many regions of the human genome contain gene families that often include pseudogenes. Other regions contain stret ...