Reverse hybridization

We have developed a simple method of nonisotopically labeling sample nucleic acids, which are then hybridized simultaneously to an array of unlabeled, immobilized probes. This "reversed hybridization" procedure thus provides identification results after a single hybridization reaction. 

Thiokol Chemical Corp, Reaction Motors Div, Study of Reverse Hybrid Propulsion System, Rept RMD 5021, QTSR-1 (Oct to Dec 1962) [Contract N600(19)-59314] (Conf)... 

The LiPA HIV-1 RT typing test is based on the reverse hybridization principle. Biotinylated DNA material obtained during PCR is hybridized with specific oligonucleotide probes immobilized as parallel lines on membrane-based strips. After hybridization, streptavidin labeled with alkaline phosphatase is added and bound to any biotinylated hybrid previously formed. Incubation with BCIP/NBT chromogen results in a purple/brown precipitate. 

Zeuzem S, Ruster B, Lee J, Stripf T, Roth W. Evaluation of a new reverse hybridization assay for genotyp-ing of hepatitis C virus. J Hepatol 1995 23 654-61. Zhang F, Ginocchio CC, Malhotra A, et al. Clinical performance and financial impact of rapid enterovirus detection in cerebrospinal fluid (CSF) using the NucliSens Basic kit and nucleic acid sequence based amphfication [Abstract T5]. 18th Annual Meeting of the Pan American Society for Clinical Virology, Clearwater, Fla, 2002. 

In reverse hybridization, unlabeled probes are immobilized on nitrocellulose membranes (0.5 xg per spot), the test sample is photo-chemically labeled (Section 7.8.2) and hybridized simultaneously to the panel of probes (Dattagupta et al., 1989). It is not necessary to purify sample DNA before or after labeling and a 300 xl sample is added to 1.1 ml of the prewarmed hybridization solution (3 X SSC, 20 mM pyrophosphate, 5% BLOTTO, 10% PEG, 1 mg/ml carDNA prehybridization 30 min, hybridization 2 h). The haptens are then detected with labeled antibody or streptavidin. 

Hybrids may be reversible or irreversible. Reversible hybrids act like sponges, soaking up the hydrogen. They are usually solids. These alloys or intermetallic compounds release hydrogen at specific pressures and temperatures. They may be replenished by adding pure hydrogen. 

Inversion of configuration (Section 8 4) Reversal of the three dimensional arrangement of the four bonds to sp hybridized carbon The representation shown illustrates inversion of configuration in a nucleophilic substitution where LG is the leaving group and Nu is the nucleophile... 

Mass spectrometer configuration. Multianalyzer instruments should be named for the analyzers in the sequence in which they are traversed by the ion beam, where B is a magnetic analyzer, E is an electrostatic analyzer, Q is a quadrupole analyzer, TOP is a time-of-flight analyzer, and ICR is an ion cyclotron resonance analyzer. For example BE mass spectrometer (reversed-geometry double-focusing instrument), BQ mass spectrometer (hybrid sector and quadrupole instrument), EBQ (double-focusing instrument followed by a quadrupole). 

The combined hybrid systems met criteria for reuse in the textile industry. Differences in performance are discussed The high-quality of the reverse osmosis permeate meets the demands of the rinsing processes for industrial laundries... 

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