Extraction method of total RNA of intermuscular bone of megalobrama amblycephala

An extraction method and technique for intermuscular bone, which can be used in DNA preparation, recombinant DNA technology, etc., and can solve the problems of inability to support molecular experiments, the amount of total RNA in intermuscular bone is low, and the extraction of total RNA has not yet occurred.

Inactive Publication Date: 2014-03-19
HUAZHONG AGRI UNIV
View PDF7 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method of extracting total RNA from the intermuscular bone which is small in size, easy to degrade and easily contaminated by muscle protein has not yet appear

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Extraction method of total RNA of intermuscular bone of megalobrama amblycephala
  • Extraction method of total RNA of intermuscular bone of megalobrama amblycephala

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A method for extracting total RNA from the intermuscular bone of bream, the steps of which are:

[0022] 1) Sampling, using bream as the material: randomly select three bream sample fish of 200-300g, in order to prevent the blood from polluting the intermuscular bone, use a sterile syringe to draw 1.5-2.0ml of blood from the tail vein of bream 1-2 minutes after death, separate the intermuscular bone from the muscle of the fish and remove the muscle tissue, immediately place it in a sterilized pre-cooled mortar, and grind in the sterilized pre-cooled mortar successively while adding liquid nitrogen, Grind to powder. Take 50mg of intermuscular bone powder from each fish, put them into 2ml sterilized centrifuge tubes pre-filled with 1ml Trizol, shake well, and place at room temperature for 4 or 5 or 6 minutes;

[0023] 2) Centrifuge the placed centrifuge tube at 4°C and 12000rpm for 5min, transfer the supernatant to another 2ml sterilized centrifuge tube, add 1 / 5 volume o...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for extracting total RNA of the intermuscular bone of megalobrama amblycephala, which comprises the following steps: 1) with megalobrama amblycephala as the material, fetching the intermuscular bone thereof and putting into a sterilized and pre-cooled mortar, and grinding until the intermuscular bone is ground into powder; transferring the powder into a centrifugal tube; adding Trizol and shaking uniformly; and placing at room temperature; 2) centrifuging at 4 DEG C, fetching the supernate, adding chloroform the volume of which is 1/5 that of the supernate, shaking so that the mixture is sufficiently emulsified into a uniform white emulsion without layering, and standing at room temperature; 3) centrifuging at 4 DEG C, fetching the supernate and repeatedly extracting with chloroform; 4) fetching the supernate, adding isometric isopropyl alcohol, sodium citrate and sodium chloride and mixing uniformly, and standing at -20 DEG C; 5) after the standing, centrifuging at 4 DEG C, discarding the supernate and leaving the precipitate, adding ethanol prepared by DEPC water, shaking and repeatedly washing and precipitating; and 6) centrifuging at 4 DEG C, discarding the supernate and leaving the precipitate, naturally airing, and adding DEPC water to dissolve RNA to obtain the total RNA solution. The process is simple, the cost is low, the obtained total RNA has high concentration, high purity and high integrity, and further molecular biology study is met.

Description

technical field [0001] The invention relates to a method for extracting total RNA from fish bones, in particular to a method suitable for extracting total RNA from intermuscular bones of bream. This method is also applicable to the extraction of total RNA from the intermuscular bone of other fishes, and the extracted total RNA can be used for cDNA synthesis and gene expression research in further molecular biology. Background technique [0002] Fish meat is more and more popular because of its high nutritional content, low fat content, easy digestion, and is suitable for the general population. Dory ( Megalobrama amblycephala ) belongs to Cyprinidae ( Cypriniformes ), Cyprinidae ( Cyprinidae ), Subfamily Cichlidae ( Culterinae ), Bream ( Megalobrama ). It is one of the important herbivorous economic fishes unique to my country. Because of its wide feeding habits, low cost, fast growth, high survival rate, easy to catch, and has the advantages of delicious taste, sm...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/10
Inventor 高泽霞万世明易少奎钟嘉王卫民
Owner HUAZHONG AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap