Download A Guide to Methods in the Biomedical Sciences by Ronald B. Corley PDF

By Ronald B. Corley

A consultant to equipment within the Biomedical Sciences supplies a simple description of universal tools utilized in learn. this isn't meant to be a tools ebook. really, it really is meant to be a ebook that outlines the aim of the equipment defined, their boundaries and supply substitute methods as acceptable. hundreds of thousands of equipment were built within the numerous biomedical disciplines and people coated during this e-book symbolize the elemental, crucial and most generally used tools in numerous assorted disciplines.

The ancient history (including a few fascinating anecdotes) resulting in the advance of ground-breaking concepts are defined, in particular those who considerably complicated the sector of biomedical learn. Advances that earned their inventors prestigious Nobel Prizes are emphasized.

The e-book is split into six sections, highlighting chosen equipment in protein chemistry, nucleic acids, recombinant DNA know-how (including forensic established methods), antibody-based concepts, microscopy and imaging, and using animals in biomedical sciences.

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When the appropriate sequence is recognized, the restriction enzymes cut the DNA at specific sites within this recognition site, giving rise to a restriction fragment, which represents the piece(s) of DNA produced after cleaving the DNA with one or more restriction enzymes. The actual cuts produced by a restriction enzyme can be “blunt” (that is, they cut at the same place on both strands of DNA) whereas others make symmetrical cuts with “overhangs”. For example, the restriction 30 A GUIDE TO METHODS IN THE BIOMEDICAL SCIENCES enzyme EcoRI (isolated from the bacterium Escherichia coli) recognizes the DNA base sequence GAATTC and cuts between the G and A (on the opposite strand the sequence is CTTAAG, and the cut occurs between the A and G on that strand).

The development of many new technologies was required to achieve these breakthroughs, and many of the techniques that were invented for these discoveries remain in use today. Some of these were so instrumental in their applicability to solve biological problems that they also garnered Nobel Prizes for their inventors (Table 2). The following are brief descriptions of some of the common methods used to manipulate nucleic acids, especially DNA. B. Basic methods for nucleic acid analysis Gels and gradients for separation of nucleic acids The size of a piece of DNA, usually produced as the result of restriction digestion (see below) can be determined by separation on gradients or gels.

The probe will be digested at any point where the probe and RNA do not hybridize. This can be the end of the RNA, or regions marking exon-intron boundaries. The remaining RNA which was “protected” by annealing to the RNA of interest is then fractionated on a sequencing gel, and the exact size and abundance of the probe remaining can therefore be determined. The size of the protected fragment reveals the start site or domain boundary, Detection and Analysis of Nucleic Acids 37 while the abundance of the protected fragment reveals the abundance of the RNA transcript.

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