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A Nature Research Journal. IN most areas of research concerned with the structure, function and synthesis of nucleic acids the measurement of molecular weights plays an inescapable part. There are many instances of problems hinging on a rather precise knowledge of the molecular weight of a nucleic acid; examples of particular current interest concern both animal and viral messengers, which are larger than can be accounted for by the total protein message that they encode, and ribosomal-RNA, the molecular weight of which bears on the question of the stoichiometry of ribosomal components.
Unfortunately the classical methods of molecular weight determination present great difficulties. The quantities of material required are frequently outside the range of feasibility. Moreover, the non-ideality in these highly charged molecules, and polydispersity 1,2 resulting from contamination with minor cellular components, or commonly products of nuclease degradation as well as the need to determine the partial specific volume , combine to make molecular weight measurement a major research undertaking.
A viscosity model of polyacrylamide gel electrophoresis. - Semantic Scholar
The remarkable spread of published values for widely studied RNA species reflects these problems. We are sorry, but there is no personal subscription option available for your country. Tanford, C. Fujita, H. Richards, E. McPhie, P. Hadjiolov, A.
Loening, U. Bishop, D. Dingman, C. Groot, P. Grivell, L.
Fisher, M. Synge , A.
- Molecular Weight Determination of Nucleic Acids by Gel Electrophoresis in Non-aqueous Solution;
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A viscosity model of polyacrylamide gel electrophoresis.
Electrophoresis and isoelectric focusing in polyacrylamide gel : advances of methods and theories, biochemical and clinical applications Richard C Allen , Hans Rainer Maurer. Richards , R. In recent years, newer techniques have emerged that give greater specificity and detail about what is happening in living systems. While these have not supplanted electrophoresis techniques, and advanced manipulations can expand the viability of the technique, it is important to realize what gel electrophoresis can and cannot do.
Electrophoresis is specific to whatever tissue you've sampled. For instance, if you run a Southern blot a type of electrophoresis on a cheek swab, you're looking at genes from the epithelial cells of your cheek and nowhere else in your body. At times, this can be beneficial, but researchers are frequently interested in more widespread effects.
Techniques such as in situ hybridization ISH can take a section of tissue and analyze gene expression at each small area of that sample. Thus, researchers can look at every brain area in a sample with ISH, whereas electrophoresis techniques can only look at a few areas at a time. Gel electrophoresis can effectively separate similar proteins with different weights this is a technique called Western blotting.
It can separate them more precisely through a technique known as 2d electrophoresis; this is common in proteomics. Unfortunately, all of the measurements made from this technique are semi-quantitative at best. In order to obtain the precise mass weight of proteins, mass spectroscopy must be employed after the protein has been purified by electrophoresis.