Oscheius tipulae nematode cryopreservation solution and preparation method and application thereof
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A technology for cryopreservation and nematodes, applied in the field of cryopreservation, can solve problems such as the inability to cryopreserve Oscheiustipulae nematodes, and achieve the effect of ensuring sustainability
Inactive Publication Date: 2014-07-02
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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[0003] The present invention will solve the problem that conventional nematode glycerol freezing solution cannot freeze and preserve Oscheius tipulae nematode, and provide a kind of Oscheius tipulae nematode freezing solution and its preparation method and the application of the freezing solution
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specific Embodiment approach 1
[0018] Specific embodiment 1: The cryopreservation solution of Oscheius tipulae nematode in this embodiment is composed of 5% to 15% agar gel, 10% to 20% tetrahydrofuran and the balance as a buffer solution according to the volume percentage;
[0019] KH in buffer 2 PO 4 The concentration is 0.003g / mL, Na 2 HPO 4 The concentration of is 0.006g / mL, the concentration of NaCl is 0.004g / mL~0.006g / mL, MgSO 4 The concentration is 0.001g / mL;
[0020] The preparation method of the agar gel is as follows:
[0021] a, KH 2 PO 4 , Na 2 HPO 4 , NaCl and MgSO 4 Add distilled water to prepare a buffer solution, then put it into a high-pressure steam sterilizer, control the temperature at 120 ℃, sterilize for 20 minutes, among them, the buffer solution KH 2 PO 4 The concentration is 0.003g / mL, Na 2 HPO 4 The concentration of is 0.006g / mL, the concentration of NaCl is 0.004g / mL~0.006g / mL, MgSO 4 The concentration is 0.001g / mL;
[0022] b. Add agar to the buffer prepared in step a, put it in a high-pre...
specific Embodiment approach 2
[0023] Specific embodiment two: this embodiment is different from specific embodiment one in that: an Oscheius tipulae nematode cryopreservation liquid contains 8%-11% agar gum, 12%-18% tetrahydrofuran and the balance by volume percentage For the buffer composition. Others are the same as the first embodiment.
specific Embodiment approach 3
[0024] Specific embodiment three: This embodiment is different from specific embodiment one in that a cryopreservation solution of Oscheius tipulae nematodes is composed of 6% agar gel, 11% tetrahydrofuran and the remainder as a buffer solution. Others are the same as the first embodiment.
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Abstract
The invention relates to an Oscheius tipulae nematode cryopreservation solution and a preparation method and application thereof, relates to a cryopreservation solution and a preparation method and application thereof, and solves the problem that a conventional nematode glycerinum cryopreservation solution can not freeze and preserve Oscheius tipulae nematodes. The cryopreservation solution comprises agaropectin, tetrahydrofuran and a buffer solution. The preparation method comprises the following steps: 1, preparing the buffer solution; 2, preparing the agaropectin; and 3, preparing the cryopreservation solution. The cryopreservation solution is used for cryopreserving the Oscheius tipulae nematode. The Oscheius tipulae nematode cryopreserved by adopting the cryopreservation solution normally grows and propagates after being unfrozen and revived, and thus the continuity of indoor culture of the Oscheius tipulae nematode captured in field is ensured. The cryopreservation solution is used for cryopreserving the Oscheius tipulae nematode.
Description
Technical field [0001] The invention relates to a cryopreservation liquid, a preparation method thereof, and application of the cryopreservation liquid. Background technique [0002] Oscheius tipulae is a nematode of the family Oscheius tipulae, which has a similar ecological niche and similar life form to C. elegans. The inventor captured Oscheius tipulae in the wild and successfully cultivated it indoors. Most nematodes can be stored for a long time at -80℃ in cryopreservation liquid with glycerin as the main component, and can grow and reproduce normally after recovery at room temperature. However, Oscheius tipulae cannot be recovered in the glycerin cryopreservation solution. Further studies have found that glycerin is fatal to Oscheius tipulae. Therefore, conventional glycerin cryopreservation solution is not suitable for long-term storage of Oscheius tipulae. Summary of the invention [0003] The invention aims to solve the problem that the conventional nematode glycerin ...
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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 王云彪郭泰亮
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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