Download e-book for kindle: Gene Probes: Principles and Protocols by Alex Reid (auth.), Marilena Aquino de Muro, Ralph Rapley
By Alex Reid (auth.), Marilena Aquino de Muro, Ralph Rapley (eds.)
The use of nucleic acid or gene probes as cloning or diagnostic instruments has promoted major advances in human genome mapping and within the isolation and manipulation of genes regularly. In Gene Probes, Marilena Aquino de Muro and Ralph Rapley have introduced jointly a very good choice of time-tested protocols for designing and utilizing gene probes in a wide selection of functions. The purposes lined variety from looking for particular genes within the human chromosome to the detection of microorganisms and their pollutants within the surroundings and in nutrients samples, in addition to within the prognosis of human affliction. every one state-of-the-art technique contains useful tricks and advice and is defined in step by step aspect through knowledgeable who not just sometimes makes use of the approach, yet has additionally optimized it for experimental good fortune. worthy tutorials clarify the foundations of gene probe layout, labeling, detection, aim formation, and hybridization stipulations.
accomplished and flexible, Gene Probes: ideas and Protocols presents a state of the art functional source for all biochemists, microbiologists, and molecular biologists who're utilizing this robust new expertise to strengthen scientific, environmental, and nutrition learn today.
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Extra info for Gene Probes: Principles and Protocols
The isotope should be stored at –20°C and should be kept behind acrylic shields at all times. [a-32P]dCTP can be used for up to 2 wk. Special care needs to be practiced during probe labeling, purification, hybridization, and washes. (II) [a-33P]dCTP (Amersham Life Science, Arlington Heights, IL). This isotope is an excellent alternative for the multiplex PCR assay. The signal is strong enough when used in PCR reactions run in thin polyacrylamide gels, and is safer to handle. Dupont-NEN also offers a stable reagent, which includes a dye (Easytides, cat.
Sterilize by autoclaving. 8. 2. Wear a mask while preparing this solution. 9. Prehybridization solution: Hybrisol I is a ready-to-use mix available from Oncor that contains 50% formamide, 6× SSC, 10% dextran sulfate, 1% SDS, sheared DNA, and modified Denhard’s solution (cat. no. S4040). 10. Membranes: High-stringency positively charged nylon membranes. 5 cm) Sure Blot Nylon Hybridization Membrane provided by Oncor (Oncor, Gaithersburg, MD), but nylon membranes provided by other vendors could also be used.
Stainsby, F. , and Bingley, M. 42 Hugenholtz, Tyson, Blackall (2000) Quantitative use of fluorescent in situ hybridization to examine relationships between mycolic acid-containing actinomycetes and foaming in activated sludge plants. Appl. Environ. Microbiol. 66, 1158–1166. 18. , and Wagner, M. (1999) The domain-specific probe EUB338 is insufficient for the detection of all Bacteria: development and evaluation of a more comprehensive probe set. Syst. Appl. Microbiol. 22, 434–444. INK4A and INK4B Alterations in Human Bladder Tumors 43 7 Evaluation of Alterations in the Tumor Suppressor Genes INK4A and INK4B in Human Bladder Tumors Irene Orlow and Carlos Cordon-Cardo 1.
Gene Probes: Principles and Protocols by Alex Reid (auth.), Marilena Aquino de Muro, Ralph Rapley (eds.)