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    Home > Biochemistry News > Biotechnology News > Gel lithography (gel draination analysis, molecular sieve layering, gel filtration, gel osmosis).

    Gel lithography (gel draination analysis, molecular sieve layering, gel filtration, gel osmosis).

    • Last Update: 2020-11-03
    • Source: Internet
    • Author: User
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    Introduction
    Gel platoonography is also known as gel exhaust chromatography,
    molecule
    sieve chromatography, gel
    filtration
    , gel filter permeation chromatography, etc. It is a liquid phase
    chromatography
    method for separating the components in the sample in molecular size order with the porous
    gel
    filler
    as the fixed phase. In 1959, Porath and Flodin first used a porous polymer-cross-linked glucosin gel as a column filler to separate samples of different molecular weights in the aqueous solution, called gel filtration. In 1964, Moore developed a cross-linked polystyrene gel with different apertures capable of separating
    organic
    solvents, called gel osmosis (gel layering with flow phases for organic solvents is commonly referred to as gel osmosis). Subsequently, this technology has been continuously improved and developed, is widely used in
    biochemistry,
    , polymer chemistry and many other fields.
    gel lithography is a commonly used separation method in biochemistry, it has the advantages of simple equipment, easy operation, high sample recovery rate, good experimental repeatability, especially does not change the biological activity of samples, so it is widely used in
    protein
    (including enzymes),
    nucleic acid
    , polysaccharides and other biological
    molecules
    separation purification, but also used in protein molecular weight determination, desalination, sample concentration and so on.

    principle of gel
    gel is to separate and purification according to the physical properties of molecular size. The layering process is shown in Figure 2-23. The fixed phase of gel layering is an inert beaded gel particle, which has a three-dimensional mesh structure inside, forming many holes. When samples containing components of different molecular sizes enter the gel
    cotography column
    , each component spreads into the hole of the fixed phase, and the degree of diffusion of the components depends on the size of the hole and the size of the component molecule. Molecules with larger aperture than the hole can not spread to the inside of the hole, completely blocked outside the hole, only in the space outside the gel particles with the flow phase downward flow, they go through a short process, flow speed, so first flow out; Long, slow flow, so the final outflow, and the molecular size between the two molecules in the flow of partial penetration, the degree of penetration depends on the size of their molecules, so they flow out of the time between the two, the larger the components of the molecules flow out first, the smaller the components flow out later. Thus, after the sample has been analyzed by gel, the components flow out in order of molecules from large to small, thus achieving the purpose of separation.









    7.2.3 Basic concepts of gel lithography
    (1) outer water volume, inner water volume, substitline volume, column bed volume, extrusion volume

    as shown in Fig. 2-24.
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