By Duckchul Park (auth.), Elena Hilario, John Mackay (eds.)
Protocols for Nucleic Acid research by means of Non-radioactive Probes, moment version presents an organization heritage at the uncomplicated preparative protocols required for the research of nucleic acids by means of nonradioactive tools. providing the methodologies utilizing extraordinary new functions, this quantity deals advisor chapters on nucleic acid extractions, instruction of nucleic acid blots, and labeling of nucleic acids with nonradioactive haptens. New fluorescent ideas equivalent to genuine Time PCR and microarrays also are incorporated, permitting clients to get a nonradioactive protocol carried out within the laboratory with minimal version required and quickest time to results.
The protocols stick with the profitable equipment in Molecular Biology™ sequence layout, each one providing step by step laboratory directions, an advent outlining the rules in the back of the procedure, lists of the mandatory gear and reagents, and tips about troubleshooting and heading off recognized pitfalls.
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Miniblotter® (MN-45, Immunetics; Fig. 3B). Manifold (F2, Immunetics; Fig. 3B). Plastic cushions (PC2, Immunetics). Nylon membranes, positively charged (Roche Applied Science). , Lumi-Film, Roche Applied Science). Alkaline phosphatase (AP)-linked antibody (Anti-DIG–AP, Fab fragments, Roche Applied Science). CDP-Star (Roche Applied Science). UV crosslinker. Hybridization oven/shaker. Flat-ended forceps. Reciprocating shaker. Heat sealer. , water pump). 72-µm Homogenizer bags (Guest Medical Ltd, Edinbridge, Kent, UK).
Eds. (1994) Current Protocols in Molecular Biology. John Wiley and Sons, New York. 2. Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual. , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. 3. Reid, A. (2002) Target format and hybridization conditions, in Gene Probes, Principles and Protocols. (Aquino de Muro, M. and Rapley, R. ), Humana, Totowa, NJ, pp. 1–11. 4. Dubitsky, A. (1997) Blocking strategies for nylon membranes used in enzymelinked immunosorbent assays.
Take care to sample the tissue in which you are interested in a representative manner. Tissue samples can be added directly to preweighed Eppendorf tubes or storage containers or to homemade tinfoil pouches immersed in the larger liquid nitrogen container. If using Eppendorf tubes, prepare the tubes with a small hole (heat a needle over a flame and pierce the lid) to prevent explosions because of the remnants of liquid nitrogen inside the sealed tube when the lids are closed. This can also be done with other containers, or else the container can be drained before placing the lid on the container.