First, the basic principle A certain amount of microorganisms, inoculated in a suitable fresh liquid medium, cultured at a suitable temperature, the logarithm of the number of bacteria as the ordinate, the growth time as the abscissa, the curve is called the growth curve. Generally, it can be divided into four periods: delay period, log phase, stable period and decay period. Different microorganisms have different growth curves, and the growth curves of the same microorganism under different culture conditions are different. Therefore, measuring the growth curve of a microorganism is very helpful for understanding and mastering the growth pattern of microorganisms.
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There are many methods for measuring the growth curve of microorganisms, such as blood cell counting method, plate colony counting method, weighing method, and turbidimetry. In this experiment, the turbidimetric method is used. Since the concentration of the bacterial suspension is proportional to the turbidity, the optical density of the bacterial suspension can be measured by a photoelectric colorimeter to infer the concentration of the bacterial liquid, and the measured optical density. The value (OD value) is plotted against its corresponding culture time, and the growth curve of the bacteria under certain conditions can be plotted. Photoelectric colorimeter (Fig. VII-10), which can directly measure the OD value by using a test tube, can be made by inoculation of a test tube and periodically measuring it.
2. The equipment is cultured for 18-20 hours of E. coli culture medium, and 12 large test tubes containing 5 ml of meat paste peptone liquid medium;
Type 72 or 72.1 spectrophotometer, self-controlled water bath shaker or shaker, sterile pipette, etc.
Third, the operation steps 1. The numbered 11 large test tubes containing the meat paste peptone liquid medium were marked with a marker to indicate the incubation time, ie 0, 1.5, 3, 4, 6, 8, 10, 12, 14, 16, 20 hours.
2. Inoculate with a 1 ml sterile pipette, and accurately inoculate 0.2 ml of E. coli culture solution each time, inoculate the numbered 11 pieces of meat paste peptone liquid medium in a large test tube, inoculate and inoculate, and mix the cells.
3. Culture The 11 tubes after inoculation were placed on a self-controlled water bath shaker or shaker and shake cultured at 37 °C. The test tubes numbered as corresponding time were taken out at 0, 1.5, 3, 4, 6, 8, 10, 12, 14, 16, and 20 hours, and immediately stored in the refrigerator, and the optical density value was determined by turbidity after a year. .
4. The turbidimetric assay was performed using a non-inoculated meat paste peptone medium as a blank control, and a photoelectric turbidity measurement was performed using a wavelength of 540-560 nm. The test is carried out sequentially from the bacterial suspension of zui concentration, and the bacterial suspension having a large concentration is appropriately diluted with the uninoculated meat paste peptone liquid medium, and the optical density value is within 0.1-0.65. When recording the OD value, take care to multiply the diluted multiple.
Fourth, the experimental report