Basic technology of biochemical experiment: crushing technology.

In addition to certain cells,
peptide
hormones
and certain
proteins
and enzymes, for in-cell or multicellular organisms
tissue
in the separation and purification of various biomolecules, all need to be broken in advance, so that the biometrics fully released into the solution, without loss of biological activity. Different organisms or different parts of the same organism of the tissue, its cell fragmentation is difficult to vary, the use of different methods, such as animal organs of the cell membrane is more fragile, easy to break, plants and
microorganisms

1. Mechanical Crushing
(1) Grinding
Grinding is the use of sheared animal tissue in a research or homogenizer, adding a small amount of quartz sand grinding or homogeneity, you can break animal cells, this method is relatively mild, suitable for laboratory use. Electric grinding is available in industrial production. This method can also be used for the fragmentation of bacterial and plant tissue cells.
(2) homogeneity
is a more intense method of breaking cells, usually using a household food processor to break the tissue, and then with 10000r/min-20000r /min's inner knife tissue masher (i.e. high-speed dispersor) breaks the tissue cells, usually turning 10 seconds-20 seconds, stopping 10 seconds-20 seconds, and can be repeated multiple times to prevent heating and heating up too high.
(3) Collate grinding
the basic principle of collory grinding is that fluid or semi-fluid material under the influence of centrifugal force, forced through high-speed relative movement between the fixed teeth and moving teeth, so that the material by a strong shear force, friction force, high-frequency vibration and high-speed vortex and other complex forces, effectively crushed, emulsified, homogeneous, mixed, so as to achieve a fine super-fine effect. The high-speed relative motion between the fixed rotor is the main condition for the collectant grinding work to obtain physical fineness, and only by improving the precision and line speed of the grinding disc can we achieve good processing results.
2. Physical
(1)
is a gentle, thoroughly broken cell method. At 1000×105Pa-2000×105Pa at high pressure so that dozens of milliliters of cell suspension through a small hole suddenly released to normal pressure, the cells will be completely broken. This is an ideal method for breaking cells, but the instrument costs more.
(2) hot and cold alternate
hot and cold alternate is maintained at about 90 degrees C for a few minutes, immediately put in an ice bath to cool it, so repeatedly, the vast majority of cells can be broken, bacteria or viruses to extract proteins and
nucleic acid
when this technique can be used.
(3) Repeated freezing
repeated freezing is to cool the cells to be broken to 15 degrees C-20 degrees C, and then put at room temperature (or 40 degrees C) to melt rapidly, so repeatedly freeze and thaw many times, due to the formation of ice particles in the cells to increase the salt concentration of the remaining cell fluid caused by cell fragmentation.
(4) ultrasound
ultrasound is the vibrational force of ultrasonic vibration to break cell walls and cells. When breaking microbial bacteria
and
yeast bacteria, the time is longer, and the effect of treatment is related to sample concentration and frequency of use. Take care to cool down when using to prevent overheating.
.