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Title: Cell Lysis Techniques
Description: These notes have complete detail about techniques used in lysis of cell . Cell lysis is important because of its of uses; it is used in western and Southern blot, lipids, nucleic acids , protein purification, reporter assays, and immunoassays etc.
Description: These notes have complete detail about techniques used in lysis of cell . Cell lysis is important because of its of uses; it is used in western and Southern blot, lipids, nucleic acids , protein purification, reporter assays, and immunoassays etc.
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Cell lysis
LYSIS:
Lysis (Greek lýsis from lýein "to separate")
refers to the breaking down of a cell, often by
viral, enzymic, or osmotic mechanisms that
compromise its integrity
...
Definition
Cell lysis is a common outcome of viral
infection
...
Methods of Cell Lysis
There are two main methods of cell Lysis:
Mechanical method of cell Lysis
Non-mechanical of cell Lysis
Mechanical Methods of Cell
Lysis
Mechanical cell disruption is really just that: forcing
open the cell wall and spilling the contents
...
The drawback however is that the method has to be
carefully adjusted as to not destroy that molecule of
interest
...
This does not
have to be in suspension and is often done with plant
samples frozen in liquid nitrogen
...
Beadbeating
Glass or ceramic beads are used to crack open cells –
it might not sound like it, but this kind of mechanical
shear is gentle enough to keep organelles intact
...
A process called cavitation occurs,
in which tiny bubbles are formed and explode,
producing a local shockwave and disrupting cell
walls by pressure change
...
Homogenizer
Homogenizers use shearing forces on the cell similar
to the bead method
...
Freezing:
Freeze-thaw cycles work by formation of ice crystals
and cell expansion upon thawing, ultimately leading
to rupture
...
The drawback is that it is very time-consuming
...
Although it is quick, you better find another method
if your application is affected by the damage heat
does to the rest of the cell
...
They are often used in
combination with mechanical force to ensure
complete disruption of the cell
...
Enzymes:
Naturally occurring enzymes can be used to remove
the cell wall specifically, for example when isolating
the protoplast (cell without the wall)
...
Chemicals:
Organic solvents like alcohols, ether or chloroform
can disrupt the cell wall by permeabilizing cell
walls and membranes
...
Often used on plants in combination with
shearing forces
...
EDTA will chelate the
cations leaving holes in the cell walls
...
Because SDS is similar in
structure and polarity to phospholipids, it can
disrupt and dissolve the phospholipid membrane
...
) And
dissolving the cell's membrane lyses the cell
...
Cell Lysis is used in laboratories to break open cells
and purify or further study their contents
...
Title: Cell Lysis Techniques
Description: These notes have complete detail about techniques used in lysis of cell . Cell lysis is important because of its of uses; it is used in western and Southern blot, lipids, nucleic acids , protein purification, reporter assays, and immunoassays etc.
Description: These notes have complete detail about techniques used in lysis of cell . Cell lysis is important because of its of uses; it is used in western and Southern blot, lipids, nucleic acids , protein purification, reporter assays, and immunoassays etc.