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A kind of preparation method and application of glass micropipette

A pipette, micro-volume technology, applied in the field of micromanipulation and molecular biology, to achieve the effect of simple method and simple collection of materials

Active Publication Date: 2017-10-17
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no suitable method to ensure the separation of high-purity liquid endosperm and the extraction of high-concentration and high-quality DNA. This application establishes a technology for quickly and efficiently separating high-purity liquid endosperm and an improved liquid endosperm genomic DNA extraction technology. The development of the subsequent endosperm and the regulation mechanism of the interaction between the endosperm and the embryo laid the foundation

Method used

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  • A kind of preparation method and application of glass micropipette
  • A kind of preparation method and application of glass micropipette
  • A kind of preparation method and application of glass micropipette

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Experimental program
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Embodiment 1

[0038] A preparation method of a glass micropipette, comprising:

[0039] The 1mm glass capillary is heated on the flame of an alcohol lamp. After the heated area of ​​the capillary becomes soft, it is slowly pulled away from both ends of the capillary. The capillary finally breaks and forms a very thin tip. The formed tip is polished to form an opening with a diameter of 25-30 μm. One end of the tip is connected with a rubber hose, and the end of the rubber hose is sealed with a glass stopper to obtain a glass micropipette ( figure 1 ).

[0040] The glass stopper is capillary fired.

Embodiment 2

[0042] Determining the stage of development of ovules for isolation of liquid endosperm

[0043]In this example, the flowers of Brassica napus ZY036 that are blooming are marked with a thin line. Subsequently, the siliques of 8 days, 16 days, 22 days, 24 days, 25 days, 26 days and 28 days after flowering were taken respectively for observation, and it was found that the length of the siliques was approximately finalized in 25 days after flowering ( figure 2 Middle A). In addition, the ovules of different time periods were separated under a dissecting microscope, and then observed under a stereomicroscope, it was found that the volume of the ovules changed the fastest from 8 days to 22 days after flowering, and the volume of the ovules was basically finalized on the 25th day ( figure 2 Middle B). Therefore, the ovules 25 days after flowering were finally selected as the material for separating the endosperm, which was used in the following examples.

Embodiment 3

[0045] Selection of the Best Perforation Position on the Ovule Surface of Brassica napus ZY036

[0046] The embryos in ZY036 25DAF ovules were separated under an inverted microscope (OLYMPUS IX71) by stripping needles, and their morphology was observed. It was clear that the embryos of 25DAF ZY036 were torpedo stage embryos (torpedo stage) ( image 3 Middle A); according to the size and morphological structure characteristics of ZY03625DAF ovules and embryos, the position and occupied space of 25DAF embryos in the whole ovule were simulated by Photoshop ( image 3 Medium B and image 3 Middle C), it is clear that on the surface of the ovule far away from the embryo and close to the chalazal end is the best reasonable position for punching ( image 3 Above the black dotted line shown in B, near the chalazal point is the perforating safe area).

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Abstract

The invention discloses a preparation method and application of a glass micro-liquid sucking device. The preparation method comprises the following steps: heating a 1-mm glass capillary tube until the tube becomes soft; then drawing two ends of the tube; polishing a sharp end formed after fracture into an opening with a diameter of 25 to 30 [mu]m; connecting a non-sharp end to a flexible rubber pipe; and sealing the tail end of the flexible rubber pipe so as to obtain the glass micro-liquid sucking device. The glass micro-liquid sucking device can realize micro-separation of trace liquid tissue. The glass micro-liquid sucking device is used for separating high-purity liquid endosperm of cabbage type rape in the invention. The invention also discloses an extraction method for the genomic DNA of the liquid endosperm, so a high-purity material and DNA are provided for research on the growth of triploid endosperm and the mechanism of interaction between endosperm and an embryo.

Description

technical field [0001] The invention belongs to the technical fields of micromanipulation and molecular biology. More specifically, it relates to a preparation method and application of a glass micropipette. Background technique [0002] Rapeseed is the third largest oil crop in the world and the fifth largest crop in my country after rice, wheat, corn and soybean. In addition to being the main raw material for edible oil, rapeseed is also an important industrial raw material and a major biological resource for renewable energy in the main member states of the European Union. The planting area and total output of rapeseed in my country account for about 30% of the world's total. It is not only related to the adjustment and optimization of my country's planting industry structure and the income increase of nearly 100 million farmers, but also related to the improvement of people's living standards, improvement of dietary structure and national security. The need for food saf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01L3/02C03B23/047C12N5/04C12N15/10
CPCB01L3/021C03B23/047C12N5/04C12N15/1003
Inventor 华玮李俊范世航孙兴超胡志勇刘静朱晓义郑明孙凤明
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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