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High-throughput screening method of oil-rich microalgae

A screening method and high-throughput technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high-throughput determination of oil content, toxic reagents, human and environmental hazards, time-consuming and labor-intensive problems, etc. problems, to achieve the effect of avoiding traditional oil detection steps, fast measurement and less time-consuming

Inactive Publication Date: 2013-06-12
HARBIN INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to screen the oil-rich algal species that can be mixed and cultivated, and to solve the time-consuming and labor-intensive problems of the existing microalgae organic solvent extraction method, which requires a large amount of reagents and samples, and toxic reagents will cause harm to the human body and the environment. It is difficult to meet the needs of high-throughput determination of oil content in microalgae screening, and provide a high-throughput screening method for oil-rich microalgae

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  • High-throughput screening method of oil-rich microalgae
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  • High-throughput screening method of oil-rich microalgae

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specific Embodiment approach 1

[0019] Embodiment 1: A high-throughput screening method for oil-rich microalgae in this embodiment is achieved through the following steps:

[0020] 1. Take microalgae samples and add them to medium A for enrichment and culture for 15 to 30 days to obtain algae liquid; 2. Take the algae liquid obtained in step 1 and inoculate in medium A with an inoculation amount of 10% by volume , cultured in a light incubator at a temperature of 25°C for 7 to 14 days; 3. Repeat step 2 2 to 3 times to obtain algae liquid A; 4. Take the algae liquid A obtained in step 3, and the volume percentage is Inoculate 10% of the inoculum into medium B, and cultivate in an incubator at 25°C for 7 to 14 days to obtain algae liquid B; -1 、10 -2 、10 -3 、10 -4 、10 -5 and 10 -6 Dilute the multiple of 10 -5 ~10 -6 The algae liquid after multiple dilution is evenly spread on the solid medium C plate, and cultivated in a 25°C incubator until algal colonies appear; 6. In a sterile environment, pick a sin...

specific Embodiment approach 2

[0028] Specific embodiment two: The difference between this embodiment and specific embodiment one is that the microalgae samples described in step one are Chlorella, Scenedesmus, diatoms, Cryptodinoflagellates, Flatweed, Dunaliella, Spirulina, One or several types of golden algae are mixed in any ratio. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0029] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: the culture medium A described in step one and step two is made of 1 part of sodium nitrate, 0.075 part of magnesium sulfate, 0.04 part of phosphoric acid in parts by weight Dipotassium hydrogen, 0.036 parts of calcium chloride, 0.02 parts of sodium carbonate, 0.006 parts of citric acid, 0.006 parts of ferric ammonium citrate, 0.001 parts of ethylenediaminetetraacetic acid and 0.00541 parts of trace elements. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a high-throughput screening method of oil-rich microalgae, relates to the field of microalgae biological energy, and aims at solving the problems that an existing microalgae organic solvent extraction method consumes time and labor and requires a large quantity of reagents and samples, a toxic reagent does harm to human body and environment, and the content of oil is hard to measure with high throughput during microalgae screening. The method comprises the following steps of: (1) fetching microalgae and performing enrichment culture for 7-14 days; (II) repeating for twice to three times; (III) after inoculation, culturing for 7-14 days; (IV) diluting and spraying, and culturing until algae colonies appear; (V) choosing large pure algae colonies with a high growth speed, and culturing for 7-14 days; (VI) repeating steps (IV) to (V) for 3-4 times; and (VII) performing ultrasonic breaking, adding a Nile red solution, and selecting an algae strain with the highest fluorescence intensity to obtain an algae strain with a high oil content. The method disclosed by the invention avoids complicated traditional oil detection steps, and overcomes the problem of bottleneck that a quick high-throughput screening method is unavailable in large-scale screening of algae seeds.

Description

technical field [0001] The invention relates to the field of microalgae bioenergy. Background technique [0002] In recent years, with the decreasing reserves of fossil fuels, the depletion and non-renewability of fossil fuels and the global warming and environmental pollution caused by their combustion, all countries in the world are forced to look for green and sustainable new alternatives to fossil fuels. energy. Biomass energy can use photosynthesis to store solar energy in the form of chemical energy in organisms, and is the most common environmentally friendly renewable energy on the earth. Among the many biomass energies, biodiesel has received widespread attention as the fastest growing and most widely used green new energy in the world. [0003] The main component of biodiesel is fatty acid methyl ester, which has similar performance to petrochemical diesel, and has excellent combustion performance, environmental protection and renewability. Its main raw material...

Claims

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Application Information

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IPC IPC(8): C12N1/12G01N21/64C12R1/89
Inventor 任南琪任宏宇刘冰峰谢国俊赵磊马超
Owner HARBIN INST OF TECH
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