A kind of yeast source nucleic acid feed additive and its application
A feed additive, yeast technology, applied in application, animal feed, additional food elements, etc., to reduce breeding costs, significant economic benefits, and improve animal health.
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Embodiment 1
[0043] The preparation of embodiment 1 bacterial classification
[0044] Source of the strain: the strain of Schwanniomyces occidentalis was purchased from China Food Fermentation Research Institute, China Industrial Microorganism Culture Collection Center, No. 32911.
[0045] (1) Inoculate Schwann yeast on plate medium, culture at 30° C. for 24 hours, and obtain activated cells. Plate medium composition: 3.0g yeast extract, 1.0g malt extract, 3.0g soluble starch, 5.0g peptone, 10.0g glucose, 20.0g agar, 1.0L distilled water, pH6.5. Use after autoclaving at 121°C.
[0046] (2) Seed culture: inoculate the bacteria in step (1) into the seed medium, and culture at 30° C. and 200 rpm for 24 hours to obtain seed liquid. The composition of the seed medium: 3.0g of yeast extract, 1.0g of malt extract, 3.0g of soluble starch, 5.0g of peptone, 10.0g of glucose, 1.0L of distilled water, pH6.5. It is used after autoclaving at 121°C.
[0047] (3) Fermentation culture: inoculate the seed...
Embodiment 2
[0048] DNA and RNA content determination in the wet thalline of embodiment 2
[0049] (1) The DNA extraction method is as follows:
[0050] Reagents: SE buffer (1M sorbitol, 0.1M EDTA, pH7.5, solvent is water); lysozyme (50mg / ml); 20% PVP; proteinase K buffer (10mM Tris, pH7.6, 0.5% SDS , 1 mM EDTA, the solvent is water).
[0051] Dissolve 0.2 g of the wet bacteria obtained in Example 1 in 590 μL of SE buffer and mix well, add 10 μL of lysozyme, react at 37 ° C for 1 hr, centrifuge at 12000 rpm for 8-10 min, collect the precipitate and dissolve it in 100 μL of proteinase K buffer, add Mix well in 0.5 μL proteinase K (1 mg / mL aqueous solution), react at 37°C for 1 hour, add 1.2ml of SDS / NaCl (1% SDS, 100 mM NaCl), 200 μL 20% PVP, mix well, and react at 65°C for 30 minutes. Add an equal volume of saturated phenol / chloroform / isoamyl alcohol mixture, mix well, centrifuge at 12000rpm for 4-5min, and repeat once. Take the supernatant, add ethanol for precipitation, dry, add 1 μL ...
Embodiment 3
[0061] Embodiment 3 preparation of yeast source nucleic acid feed additive
[0062] Composition of solid-state fermentation medium: bran 500g, corn flour 550g, soybean meal 500g, molasses 250g, water 1800g, pH natural. The above-mentioned raw materials can be purchased in the market.
[0063] The fermented liquid containing the wet thalline prepared in Example 1 was inoculated into the solid-state fermentation medium in an amount of 10% (the weight percent of the fermented liquid containing the thalline and the solid substrate), and fermented and cultivated at 30° C. for 48 h. Turn the material once every 12 hours. After the fermentation is over, the fermentation product is steam-dried to a moisture content of ≤11.5%, and then crushed to a fineness of 40-60 mesh to obtain 1756 g of yeast-derived nucleic acid feed additive. Sampling, RNA and DNA were extracted from the ground sample according to the method in Example 2, and the total amount of nucleic acid was calculated to be...
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