The refrigerator freezing method of microbial species preservation method -80 degrees C.

Biotechnology Channel News: -80 degrees C refrigerator freezing method to keep the bacteria in the -80 degrees C refrigerator to slow down the physiological activity of cells for freezing a method of preservation.
operation steps are as follows: 1, ampoule tube preparation ampoule tube material to neutral glass is appropriate.
When cleaning the ampoule tube, first soaked with 2% hydrochloric acid overnight, after washing clean tap water, soaked with distilled water to pH neutral, dried, labeled, labeled with bacteria number and time, added to the skimmed cotton plug, 121 degrees C autocultension 15-20 minutes, spare.
2, the choice and preparation of protective agents to be selected according to the microbial category.
preparation of protective agents, attention should be paid to their concentration and pH, as well as sterilization methods.
such as serum, can be filtered to sterilise; milk should be skimmed first, with centrifugal method to remove the upper fat, generally boiled 2-3 times at 100 degrees C intermittently, 10-30 minutes at a time, spare.
3. Preparation of microbial preservation The cells are cultured to a stationary or mature stage under the most suitable culture conditions, after purity inspection (see the Office of Joint Management of natural science and technology resources platform of the Ministry of Science and Technology document "Technical Procedures for The Testing of Microbial Species Purity" (trial)), mixed with the protective agent evenly and divided. the concentration of
microbial cultures is suitable for cells or spores not less than 108-1010 /ml (E. coli, for example, in order to obtain 1010 live cell fluid per milliliter 2 ml-2.5 ml, only 10 ml agar slope two).
using a longer capillary dropper, directly drip into the bottom of the ampoule tube, pay attention not to spill the upper tube wall, each tube assembly volume of about 0.1-0.2 ml, if it is a spherical ampoule tube, the loading capacity of half the ball.
if it is a liquid cultured microorganism, the medium should be centrifuged to remove the medium, and then the culture is mixed with the protective agent, and then divided into ampoule tubes.
as short as possible, preferably within 1-2 hours and prefrozen.
should be operated under sterile conditions when packing.
4, frozen storage The ampoule tube or plastic freezer tube is placed in the refrigerator at -80 degrees C.
5, resuscitation method Remove the ampoule tube or plastic freezer tube from the refrigerator, should immediately place 38 degrees C-40 degrees C water bath to quickly recover and shake as quickly as appropriate.
until all the internal ice dissolves, it takes about 50-100 seconds.
the ampoule tube or plastic freezer tube and move the contents to a suitable culture base for culture.
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