The separation and purification of microorganisms

, the purpose requires
master the method of inverting the plate and several commonly used basic operating techniques for separating purified materials.
, the basic principles
1. The process of obtaining
containing only one
microorganisms from a mixed group of microorganisms is called the separation and purification of microorganisms. This experiment uses flat-panel separation method: the method is simple to operate, widely used in microbial separation and purification, which includes two aspects:
(1) select suitable for the growth conditions of the microorganism to be separated, etc. , or add an inhibitor to cause only the growth of the microorganism, while inhibiting the growth of other microorganisms in the environment, thereby eliminating some unwanted microorganisms;
(2) A single organism formed by microorganisms growing on solid
cultures
-based can be a collection of cells, so a pure culture can be obtained by selecting a single bacteria. The method of obtaining a single bacterios can be done by diluting counts such as coated plates or flat dashes.
2. Determination of pure culture:
(1) to determine the observation characteristics of its bacteria,
(2) to detect the determination of individual morphological characteristics in combination with
smicror microscope

third, equipment
1. Bacteria: mystic mold;
2.
culture
: Gaussi I. culture, beef paste proteinculture, Martin's
argisit
culture, cha's agar culture;
3. Solution or
reagent
:10% phenol containing 9 ml of sterile water
test tube
90 ml of sterile water with glass beads of triangular burners, 4% water agar;
4. Instruments or other utensils: sterile glass sticks, sterile straws, inoculation rings, sterile petri dishes, streptomycin and soil samples, microscopes, blood cell counting boards, etc.
, the operation steps
1. Diluted coated plate method
(1) inverted plate: add the paste proteinagar medium, Gaussi I. agar medium; Martin's agar medium
heat
dissolve, to be cooled to 55-60 degrees C, Gao's I. agar medium added 10% phenol drops, Martin's agar medium added chain mold solution. After mixing, pour the plate, each media pour three dishes;
(2) prepare soil dilution solution: called soil sample 10g, put in a triangular bottle containing 90 ml of sterile water and brought into the glass beads, vibration about 20min, so that the soil sample mixed with water, cell dispersion. 1ml1ml9ml,1ml9ml,,10-1,10-2,10-3,10-4,10-5,10-6；
（3）：10-4,10-5,10-6,10-4,10-5,10-60.1ml5-10min,；
（4）：Ⅰ; Martin's agar medium plate was cultured for 3-5 days in a greenhouse at 28 degrees C, and the meat paste protein
(5) Pick the bactericides: the cultured individual bacteria grow out of a small number of cells inoculated to the slope of the above three cultures, respectively, placed 28 degrees C and 37 degrees C greenhouse culture, bacteria moss grow out, check whether their characteristics are consistent, while the cell smear stained with a microscope to check for a single microorganism. If clutter is found, it needs to be separated and purified again until pure culture is obtained.
2. Flat dash separation method
(1) inverted plate: according to dilution coated plate inverted plate, and marked with a marker of the culture base name, soil sample number and experimental date;
(2) dash: near the flame, left hand holding the bottom of the dish, right hand holding the inoculation ring, pick the above soil suspension ring on the flat plate. There are many methods of dashing, but regardless of the method used, the purpose is to draw the sample on the plate to dilute it to form a single bacterios,
(3) pick the bacterios: the same dilution coating plate method, until the isolated microorganisms consider purification.