Add probes

Figure 36- i Purine and pyrimidine bases and the formation of complementary base pairs. Dos/ied //nes indicate the formation of hydrogen bonds. (In RNA, thymine is replaced by uracil, which differs from thymine only in its lack of the methyl group.) (Modified from Piper, M.A., Unger, E.R. Nucleic Add Probes A Primer for Pathologists. Chicago, ASG Press, 1989.)... 

Figure 37-19 Melting curve of double-helical nucleic acid. (Modified with permission from Piper MA, Unger ER Nucleic Add Probes A Primer for Potho/og/sts. Chicago, ASCP Press, 1989.)...

Li Q, Luan G, Guo Q, Liang I. A new class of homogeneous nucleic add probes based on specific displacement hybridization. Nucleic Acids Res 2002 30 E5. 

Gently heat cells to 60-70°C to denature DNA, add probe and incubate at the hybridization temperature (usually room temperature). 

Remove prehybridization solution without permitting the specimens to dry and add probe in hybridization solution (radiolabeled probes 10—600 ng/ml nonradioactive probes up to 10 times more). Hybridization solution contains 50% formamide, 0.5 M NaCl, 10 mM Tris-HCI (pH 7.6), 2 mM EDTA, 1 x Denhardt s solution, 20 mM DTT, 300 pg/ml tRNA, 300 (xg/ml poly(A) or/and poly(C) (optional), 10% PEG 6000 RNase-free. S label may form nonspecific disulfide bonds in the cell if reductants, such as DTT or 2-mercaptoethanol, are omitted. 

J. Wang, M. Jiang, A. Fortes, and B. Mukherjee, New label-free DNA recognition based on dicing nucleic-add probes within conducting polymer films. Anal. Chim. Acta 402, 7-12 (1999). 

Incubate at 37°C for at least 15 min but up to several hours, until you are ready to add probe. The slow exchange into and preincubation in 50% formamide/ 2 X SSCT are to ensure that the embryos are completely equilibrated in this solution. 

Place slides section side up to air dry on foil. Add probe to them the same day. 

Add probe to hybridization solution (10-200 ng/slide—actual amount determined empirically). Heat to 70-80°C for 5 min to ensure uniform solution and quench in ice. 

Amann RI (1995) In situ identification of micro-organisms by whole cell hybridization with rRNA-targeted nucleic add probes. Molec Microbiol Ecol Man... 

We improve the penetration of eddy currents by increasing the probe diameter. The useful diameter is generally equal to the coil diameter to which we add four times the standard thickness 8. 

A connnon teclmique used to enliance the signal-to-noise ratio for weak modes is to inject a local oscillator field polarized parallel to the RIKE field at the detector. This local oscillator field is derived from the probe laser and will add coherently to the RIKE field [96]. The relative phase of the local oscillator and the RIKE field is an important parameter in describing the optical heterodyne detected (OHD)-RIKES spectrum. If the local oscillator at the detector is in phase with the probe wave, the heterodyne mtensity is proportional to... 

Ion probes. Determining the level of ions in solution also helps to control corrosion. An increase in concentration of specific ions can contribute to scale formation, which can lead to a corrosion-related failure. Ion-selective elec trode measurements can be included, just as pH measurements can, along with other more typical corrosion measurements. Especially in a complete monitoring system, this can add information about the effect of these ions on the material of interest at the process plant conditions. 

Watts AG, Withers SG (2004) The synthesis of some mechanistic probes for sialic add processing enzymes and the labeling of a sialidase from Trypanosoma rangeli. Can J Chem 82 1581-1588 Watts AG, Oppezzo P, Withers SG, Alzari PM, Buschiazzo A (2006) Structural and kinetic analysis of two covalent sialosyl-enzyme intermediates on Trypanosoma rangeli sialidase. J Biol Chem 281 4149-4155... 

Kidney, liver, and muscle. Weigh 25 g of sample into a 250-mL polypropylene centrifuge bottle. Make fortifications to the samples at this point. Add 100 mL of acetonitrile-water (4 1, v/v) and 10 drops of Dow Corning Antifoam B emulsion to the sample bottles, and homogenize the samples with a Polytron at high speed for 1 min. Rinse the Polytron probe with an additional 80 mL of acetonitrile-water... 

NMR microscopy systems are built as add-ons to NMR spectrometers. The general technology of an NMR spectrometer is not the subject of this chapter, only the parts that are of particular importance in NMR microscopy applications, such as the magnets, the shim systems, the gradient systems, the NMR microscopy probes and the gradient amplifiers, will be described in some detail. 

Systems have been developed by some of the major spectrometer manufacturers to deal specifically with this type of application. These systems are designed with automation very much a priority. Typically, an integrated robot adds a predetermined volume of solvent to each of the wells and then injects the resultant solution into a flow line that transfers it into the spectrometer s probe, which is of course fitted with a flow cell. Spectroscopy can then be performed without the time constraints of the HPLC-NMR system and the sample returned to the well on the plate where it came from, or into a fresh one if required. 

Add a 10-fold molar excess of AMBH (pre-dissolved in ethanol) (Molecular Probes) over the expected amounts of aldehydes to be modified. 

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