Preparation of botryococcus protoplast and method for measuring preparation rate of botryococcus protoplast
A technology of protoplasts and botrytis, applied in the biological field, can solve problems such as the great influence of human factors, the reduction of sample parallelism, and the easy death of samples, and achieve the effects of saving manpower, improving preparation rate, and improving objectivity
Inactive Publication Date: 2014-10-15
SOUTH CHINA UNIV OF TECH
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However, this method requires manual counting under a microscope, which is time-consuming, and human factors have a great influence. When the sample volume is large, it is difficult to accurately complete the calculation of the preparation rate in a short time, making the sample easy to die, and the parallelism between samples reduce
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Embodiment 1
[0040] This embodiment is an embodiment of the preparation method of botrytis protoplasts and the detection method of the production rate of botrytis protoplasts.
[0041] (1) Botryococcus sudeticus UTEX2629 (Botryococcus sudeticus UTEX2629, referred to as UTEX2629) algae were inoculated on the slant Modified Bold3N medium, the culture conditions were light / dark 12 / 12h, and the light intensity was 2000lux and 25°C for 15 days. Pick a ring of algal moss (slope seeds cultivated at 25°C) and inoculate it in a 50mL Erlenmeyer flask filled with 20mL Modified Bold3N medium, light / dark 12 / 12h, light intensity 2000lux, 25°C, 130r / min shaking culture After 10 days, the inoculum was inoculated into a 250mL Erlenmeyer flask containing 100mL of sterile Modified Bold3N medium at an inoculum size of 20% (v / v), and cultured at 25°C with a light intensity of about 4000lux and a rotation speed of 130r / min.
[0042] In this embodiment, the Modified Bold3N medium formula is: sodium nitrate (NaNO...
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The invention provides a preparation method for botryococcus protoplast and a method for measuring the preparation rate of the botryococcus protoplast. The preparation method for the botryococcus protoplast comprises the following steps: 1) selecting a single cell botryococcus UTEX2629 culture solution cultured to a mid-log phase, centrifuging and collecting alga cells, and cleaning through distilled water so as to obtain a botryococcus sample; 2) adding a high permeating compound enzyme solution into the botryococcus sample obtained in the step 1), and conducting enzymolysis under the dark condition, wherein the enzymolysis temperature is 25 DEG C, the enzymolysis time is 10-14 hours, and the compound high permeating enzyme solution comprises 0.6-0.7% (w / v) of cellulose, 0.6-0.7% (w / v) of pectinase, 1.3-1.4% (w / v) of macerozyme and 0.8 mol / L of KCl; 3) centrifuging and collecting precipitation after the enzymolysis is finished. According to the preparation method for the botryococcus protoplast, provided by the invention, the preparation rate of the botryococcus protoplast can be effectively improved, and the preparation method is simple and convenient to operate. According to the method for measuring the preparation rate, provided by the invention, the preparation rate of botryococcus protoplast by the preparation method can be quickly, simply, conveniently and accurately measured.
Description
technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for preparing botrytis protoplasts and a detection method for determining the preparation rate of botrytis protoplasts. Background technique [0002] Botryococcus is a single-celled green algae that grows in clusters. Studies have shown that botrytis algae and the hydrocarbons they produce contribute significantly to some oil shales and sediments. In fact, botrytis-specific hydrocarbons and their algal cells are present in petroleum products around the world. Hydrocarbons produced by botrytis are one of the important sources of biological hydrocarbons. They can be refined by hydrocracking, a refining procedure commonly used today for conventional petroleum. So these hydrocarbons can be easily converted into fuels and used directly in the market without the need to modify the existing fuel-dependent infrastructure and motor vehicle configuration. Therefore, botrytis has b...
Claims
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IPC IPC(8): C12N1/12C12Q1/02C12R1/89
Inventor 魏东熊爽
Owner SOUTH CHINA UNIV OF TECH