Probe preparation

Forster, A.C., Mclnnes, J.L., Skingle, D.C., and Symons, R.H. (1985) Non-radioactive hybridization probes prepared by the chemical labeling of DNA and RNA with a novel reagent, photobiotin. Nucleic Acid Res. 13, 745-761. 

Reisfeld, A., Rothenberg, J.M., Bayer, E.A., and Wilchek, M. (1987) Nonradioactive hybridization probes prepared by the reaction of biotin hydrazide with DNA. Biochem. Biophys. Res. Comm. 142, 519-526. 

Most protocols that use RNA probes recommend stringent procedures to ensure the absence, as far as possible, of contaminating RNase during the probe preparation and hybridization procedures. RNase is... 

CO2 is the gas generally used for mass-spectrometric analysis. More recently CO and O2 have also been used in high temperature conversion of organic material and in laser probe preparation techniques. A wide variety of methods have been described to liberate oxygen from the various oxygen-containing compounds. 

The use of probes prepared by a covalent attachment procedure of gold particles and proteins has been recently described as offering a number of advantages over conventional gold probes, including better resolution, stability, uniformity, sensitivity, and complete absence of aggregation (21). 

Various websites provide extensive information on gold probe preparation and use. Of particular value are the regular Nanoprobe newsletters (http //www.nanoprobes.com) and immunogold newsletters (http //www.british-biocell.co.uk). 

The analyzer operation is based on mass-spectrometric principle. The probe preparation system consists of vacuum extractor and oven. 

Add the probe (prepared as in steps 1 -4 of Procedure A) to the bag and incubate with agitation at 42° for 6-16 hr. The hybridization and washing temperatures in steps 2 and 4, respectively, should be lowered if a heterologous probe is used. When wash temperatures are below 50-55°, however, higher backgrounds may result. 

While the identification of SF3b was an impressive start, there is some concern over the considerable (645-fold) loss of activity in the Eisai probes prepared at C7. While these probes were used to identify the splicing factor SF3b, it is possible the lack in activity of Eisai s probe molecules failed to provide a complete description of the targets of the pladienolides. In addition, the mode of action of pladienolide B has yet to be directly... 

Spotting pre-synthesized DNA oligomers or DNA probes prepared in other ways ... 

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