High throughput screening method for high-yield lipase bacteria
A technology of lipase bacteria and screening method, applied in the biological field, can solve the problems of calculation result error, labor consumption, material resources, multiple disposable plates, etc., and achieve the effects of accurate results, convenient operation and material saving.
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Embodiment 1
[0025] Example 1: 96-well plate method
[0026] A high-throughput screening method was used to screen the mutagenized strains. Add 200ul of fermentation broth and a certain volume of bromophenol blue to a 96-well plate, mix it evenly, and measure its maximum absorption wavelength with an ultraviolet spectrophotometer. The maximum absorption wavelength of this solution is 570nm. According to the final result of physical radiation, single colony spores under the specific lethality induced by ultraviolet mutagenesis were selected and inserted one by one into the 96-well plate equipped with fermentation broth and bromophenol blue (3 parallel samples, wherein both sides were used as blanks) , cultivated in a constant temperature shaker at 30°C and 160r / min, and measured values with a microplate reader at different time periods. When using a microplate reader for detection, the wavelength needs to be adjusted to 570nm and preheated for 30 minutes in advance. The bacteria produce...
Embodiment 2
[0027] Embodiment 2: plate method
[0028] Pour the configured culture medium into a disposable flat plate. The average flat plate contains 5mL solution. According to the final result of physical radiation, select the single colony spores under the specific lethality induced by ultraviolet mutagenesis and insert them into the flat plate (3 cultured in a constant temperature shaker at 30°C and 160r / min, and observed regularly until the plate on which it was located was all yellow (bromocresol purple color), and the full diameters of colonies and discolorations were measured with a ruler respectively. Among them, it takes 6 days to screen by the plate method, and the final result obtained by the detection is consistent with that of the 96-well plate method.
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