By Ray Wu, Lawrence Grossman, Kivie Moldave
FROM THE PREFACE: intriguing new advancements in recombinant DNA learn let the isolation and amplification of particular genes or DNA segments from nearly any residing organism. those new advancements have revolutionized our ways to fixing advanced organic difficulties and feature spread out new probabilities for generating new and higher items within the components of wellbeing and fitness, agriculture, and industry.Volumes a hundred and a hundred and one complement Volumes sixty five and sixty eight of tools in Enzymology. over the last 3 years, many new or stronger tools on recombinant DNA or nucleic acids have seemed, and they're incorporated in those volumes. quantity a hundred covers using enzymes in recombinant DNA learn, enzymes affecting the gross morphology of DNA, proteins with really expert features performing at particular loci, new equipment for DNA isolation, hybridization, and cloning, analytical tools for gene items, and mutagenesis: in vitro and in vivo. quantity a hundred and one contains sections on new vectors for cloning genes, cloning of genes into yeast cells, and structures for tracking cloned gene expression.
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Extra resources for Recombinant DNA, Part B
Restriction enzymes isolated from thermophilic bacteria are more stable and more active at temperatures higher than 37°. 35,39,55 Selected restriction endonucleases were studied to ascertain the effect of assay temperature on their activity. EcoRI is inactive above 420, 9 while BamHI loses significant activity at 55°. , as probes of DNA secondary structure 8 or for suppression of contaminating enzymic activities. 55 Below 37° most restriction endonucleases remain active, although at reduced rates.
Compounds involved in DNA isolation should be rigorously removed by dialysis or by ethanol precipitation and drying prior to addition of the DNA sample to the restriction endonuclease reaction. For example, Hg 2+, phenol, chloroform, ethanol, ethylene(diaminetetraacetic) acid (EDTA), sodium dodecyl sulfate (SDS), and NaC1 at high levels interfere with restriction reactions, and some can alter the recognition specificity of restriction endonucleases. , actinomycin and distamycin A, 14 also influence restriction endonuclease activity.
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Recombinant DNA, Part B by Ray Wu, Lawrence Grossman, Kivie Moldave