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Silkworm egg vivisecting method

A vivisection and silkworm technology, applied in the field of silkworm egg anatomy, can solve the problems affecting the effect of nucleic acid extraction, and achieve the effect of rapid inspection, low cost, high sensitivity and convenient operation

Inactive Publication Date: 2008-10-15
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method of extracting silkworm egg nucleic acid (DNA or RNA) is to put it into liquid nitrogen together with the egg shell and then grind it. The attachment on the egg shell will contain a large number of microorganisms, which will affect the extraction of nucleic acid (DNA or RNA) Effect

Method used

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  • Silkworm egg vivisecting method

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Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1: A method for vivisection of silkworm eggs, the embryos are used for developmental analysis.

[0033] Soak the silkworm eggs in a petri dish filled with PBS buffer solution at a temperature of 50°C-55°C for 5-8 minutes, clamp and fix both sides of the silkworm eggs with tweezers, gently cut off one end of the eggs with a scalpel, and then Squeeze gently to squeeze out all the contents inside the egg. Use a plastic-tipped pipette to gently rinse the contents in water several times to separate the embryos and yolk.

[0034] Use a straw to suck out the embryo and observe it under a 400-fold microscope. According to the characteristics of the embryo, the development process of the silkworm egg can be determined.

Embodiment 2

[0035] Embodiment 2: A method for vivisection of silkworm eggs, and the embryos are used for in vivo culture.

[0036] Carry out high-temperature sterilization to all the equipment, and in the ultra-clean workbench, put the silkworm eggs sterilized by 72% ethanol for 5 minutes into a culture dish filled with sterile PBS buffer at a temperature of 50°C-55°C and soak for 5- For 8 minutes, use tweezers to clamp and fix both sides of the silkworm eggs, use a scalpel to gently cut one end of the egg, and then gently squeeze with the tweezers to squeeze out all the contents in the egg. The embryo and yolk can be separated by gently rinsing the contents in water several times with a plastic-tipped pipette.

[0037] Use a pipette to suck the complete embryos into a culture plate filled with 1mL medium (TC100), shake well, then suck out the embryos, and implant them into the culture medium for normal culture.

Embodiment 3

[0038] Embodiment 3: A kind of silkworm egg vivisection method, the embryo is used for DNA extraction

[0039] Carry out high-temperature sterilization to all the equipment, and in the ultra-clean workbench, put the silkworm eggs sterilized by 72% ethanol for 5 minutes into a culture dish filled with sterile PBS buffer at a temperature of 50°C-55°C and soak for 5- For 8 minutes, use tweezers to clamp and fix both sides of the silkworm eggs, use a scalpel to gently cut one end of the egg, and then gently squeeze with the tweezers to squeeze out all the contents in the egg. The embryo and yolk can be separated by gently rinsing the contents in water several times with a plastic-tipped pipette.

[0040] Embryos were collected, put into a mortar and ground with liquid nitrogen, and DNA was extracted according to conventional methods.

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Abstract

The invention discloses a method for vivisecting silkworm eggs, which comprises following steps: cutting one end of a silkworm egg by a scalpel, squeezing all of the content in the egg into a buffer liquid, and sucking by a sucker to separate embryo from yolk. The method is characterized in that the embryo obtained is live and has obvious characteristics to facilitate determining the development process; and the method can also be used for culture of embryo in vitro and extraction of nucleic acids (DNA or RNA). The method has important meanings for conserving resources, reducing environment pollution by strong alkali and improving labor efficiency.

Description

technical field [0001] The invention relates to a method for dissecting silkworm eggs, in particular to a method for vivisection of silkworm eggs. Background technique [0002] Silkworm (Bombyx mori L.) has been domesticated in my country for more than 5,700 years. People have been able to adjust the peripheral environment of silkworm eggs according to production needs, and control the development or diapause of silkworms at any time. It usually takes 11-12 days for silkworm eggs to hatch from diapause. During this time, people have to dissect silkworm eggs every day through traditional methods, and judge the development process of the embryos according to the developmental characteristics of the embryos in the eggs. Regulate the peripheral environmental conditions to promote uniform embryonic development. [0003] The traditional method of dissecting silkworm eggs is the alkaline method, that is, after boiling the NaOH or KOH with a concentration of 20%, immerse the silkwo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61B16/00A61B17/00
Inventor 李兵沈卫德许雅香卫正国陈玉华
Owner SUZHOU UNIV