By P. Tijssen
Fresh breakthroughs in recombinant DNA know-how and the supply of subtle apparatus available to just about any laboratory, have contributed to the improvement and perfection of robust hybridization instruments. lately, nucleic acid hybridization has not just turn into a cornerstone in molecular biology study but in addition a robust complement to different diagnostic instruments. those diagnostic equipment are set out in a logical and transparent two-part paintings during this "Laboratory recommendations" sequence. the amount is split into conception and guidance (Part I), and probe labelling and hybridization strategies (Part II).
Read or Download Hybridization with Nucleic Acid Probes: Part II: Probe Labeling and Hybridization Techniques PDF
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Additional info for Hybridization with Nucleic Acid Probes: Part II: Probe Labeling and Hybridization Techniques
Sample text
For fluorography, Kodak SB is an excellent choice due to its sensitivity for light photons. 2. The specimen is placed in contact with an X-ray film in the dark and exposed for a time which is usually a compromise between a background development and maximum signal (typically 24 h for 32P). A good contact between the specimen and film is required. ). 3. Intensifying screens are used for 32Pwhereas fluorography can be used for 35S,14C, 3H. Some highly radioactive samples may require very short exposure times or slower films without screens.
1. 035 mM fluorescein surfactant)) and use an aliquot just enough to wet the membrane. 1 M NaCl and 50 mM MgCI,) between two photocopier transparencies (acetate film, use only once), placing a few milliliters of substrate on the membrane and squeezing the liquid to cover the whole membrane. Ch. 11 (continued) * * Instead of this buffer, the rinse buffer can also be used for dilution from the stock solution. For more economical use of substrate, it may be reused but should then be stored separately in a dark bottle.
Best results are on positively charged nylon; also PVDF with Nitro-Block. 1. 035 mM fluorescein surfactant)) and use an aliquot just enough to wet the membrane. 1 M NaCl and 50 mM MgCI,) between two photocopier transparencies (acetate film, use only once), placing a few milliliters of substrate on the membrane and squeezing the liquid to cover the whole membrane. Ch. 11 (continued) * * Instead of this buffer, the rinse buffer can also be used for dilution from the stock solution. For more economical use of substrate, it may be reused but should then be stored separately in a dark bottle.