Micro-culture method of pleuromutilin producing bacteria and high-throughput screening method of high-yield bacteria of pleuromutilin
A technology of pleuromutilin and its culture method, which is applied in the field of micro-cultivation of pleuromutilin-producing bacteria and high-throughput screening of high-yielding bacteria, can solve problems such as inapplicability, and achieve improved screening efficiency, shortened fermentation cycle, The effect of reducing errors
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Embodiment 1
[0046] The microculture steps of pleuromutilin-producing bacteria are as follows:
[0047] (1) Dilute the pleuromutilin-producing bacteria 10 times with sterile water to make a bacterial suspension, spread it on a plate, incubate at 25°C for 3-5 days, and isolate a single colony;
[0048](2) Prepare a 96-well microtiter plate containing 1 / 2 of the solid fermentation medium for later use: Autoclave the solid fermentation medium at 121°C for 15 minutes, and add it to the 96-well microtiter plate in an ultra-clean bench aseptically. The amount added to the well is 150 μl;
[0049] (3) Pick up the single-colony mycelia of the pleuromutilin-producing bacteria formed in step (1) with a bamboo stick, inoculate it in a 96-well microtiter plate containing solid fermentation medium, repeat the plate once, and leave Used for identification of shake flasks;
[0050] (4) Incubate the inoculated 96-well ELISA plate at 25°C for 7 days to complete the microculture of pleuromutilin-producing...
Embodiment 2
[0065] The microculture steps of pleuromutilin-producing bacteria are as follows:
[0066] (1) Dilute the pleuromutilin-producing bacteria to 100 times with sterile water to make a bacterial suspension, spread it on a plate, incubate at 25°C for 3-5 days, and isolate a single colony;
[0067] (2) Prepare a 96-well microtiter plate containing 1 / 3 of the solid fermentation medium for later use: Autoclave the solid fermentation medium at 121°C for 15 minutes, and add it to the 96-well microtiter plate in an ultra-clean bench aseptically. The amount added to the well is 100 μl;
[0068] (3) Use an inoculation loop to pick up the single colony mycelia of the pleuromutilin-producing bacteria formed in step (1), and inoculate it in a 96-well microtiter plate containing solid fermentation medium;
[0069] (4) Incubate the inoculated 96-well ELISA plate at 25°C for 8 days to complete the microculture of pleuromutilin-producing bacteria.
[0070] The high-throughput screening method o...
Embodiment 3
[0075] The microculture steps of pleuromutilin-producing bacteria are as follows:
[0076] (1) Dilute the pleuromutilin-producing bacteria to 1000 times with sterile water to make a bacterial suspension, spread it on a plate, culture at 25°C for 3-5 days, and isolate a single colony;
[0077] (2) Prepare a 96-well microtiter plate containing 2 / 3 of the solid fermentation medium for later use: Autoclave the solid fermentation medium at 121°C for 15 minutes, and add it to the 96-well microtiter plate in an ultra-clean bench aseptically. The amount added to the well is 200 μl;
[0078] (3) Pick up the single colony mycelium of the pleuromutilin-producing bacteria formed in step (1) with an inoculation needle, and inoculate it in a 96-well microtiter plate containing solid fermentation medium;
[0079] (4) Incubate the inoculated 96-well ELISA plate at 25°C for 9 days to complete the microculture of pleuromutilin-producing bacteria.
[0080] The high-throughput screening method ...
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