Plant bleeding sap RNA extracting method

An extraction method and a technology of wounding fluid, which are applied in the field of agriculture, can solve the problems of inability to meet the needs of reverse transcription, insufficient RNA quantity, and rapid RNA degradation, so as to meet the needs of reverse transcription, increase the amount of RNA obtained, and inhibit RNA. Degradation effect

Inactive Publication Date: 2015-11-11
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, the methods for extracting RNA from plant tissues are very mature, but the effect of using these methods to extract RNA from grape, cucumber and other plant wound fluids is not sati

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
  • Plant bleeding sap RNA extracting method
  • Plant bleeding sap RNA extracting method
  • Plant bleeding sap RNA extracting method

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0023] Example 1:

[0024] A method for extracting RNA from plant wound fluid, including the following steps:

[0025] 1) Freeze-dry 5ml of plant wound fluid to 1 / 10 of the original volume, add 1400μl SDS, and water bath at 65℃ for 30min;

[0026] 2) Add 300μl of water-saturated phenol, vortex, then add 300μl of chloroform, vortex to mix, and centrifuge at 4°C, 13000rpm for 10min;

[0027] 3) Take 1200μl of supernatant to a new centrifuge tube, add 400μl of water-saturated phenol, vortex to mix, then add 400μl of chloroform, vortex to mix, centrifuge at 13000rpm for 10min;

[0028] 4) Take 1000μl of supernatant and add 500μl of water-saturated phenol, vortex to mix, then add 500μl of chloroform and vortex to mix, centrifuge at 13000rpm for 10min;

[0029] 5) Take 800μl of supernatant and add 800μl of chloroform, vortex to mix, centrifuge at 13000rpm for 10min, take 700μl of supernatant and add equal volume of isopropanol, and let stand at -20℃ for more than 5h;

[0030] 6) Centrifuge at...

Example Embodiment

[0031] Example 2

[0032] A method for extracting RNA from plant wound fluid, including the following steps:

[0033] 1) Freeze-dry the plant wound fluid to 1 / 8 of the original volume, add 1300μl SDS, and water bath at 65℃ for 25min;

[0034] 2) Add 200μl of water-saturated phenol, vortex, then add 200μl of chloroform, vortex to mix, centrifuge at 4°C, 12000rpm for 8min;

[0035] 3) Take 1100μl supernatant to a new centrifuge tube, add 1 / 3 volume of water-saturated phenol, vortex to mix, then add 1 / 3 volume of supernatant chloroform, vortex to mix, centrifuge at 12000rpm for 8min;

[0036] 4) Take 900μl of supernatant and add 1 / 2 volume of supernatant water-saturated phenol, vortex to mix, then add 1 / 2 volume of supernatant chloroform and vortex to mix, centrifuge at 12000rpm for 8min;

[0037] 5) Take 700μl of supernatant and add equal volume of chloroform, vortex to mix, centrifuge at 12000rpm for 10min, take 600μl of supernatant and add equal volume of isopropanol, and let stand at -2...

Example Embodiment

[0039] Example 3

[0040] A method for extracting RNA from plant wound fluid, including the following steps:

[0041] 1) Freeze-dry the plant wound fluid to 1 / 9 of the original volume, add 1500μl SDS, and water bath at 65℃ for 30min;

[0042] 2) Add 400μl of water-saturated phenol, vortex, then add 400μl of chloroform, vortex to mix, and centrifuge at 12,500 rpm at 4°C for 9 minutes;

[0043] 3) Take 1300μl of supernatant to a new centrifuge tube, add 1 / 3 volume of water-saturated phenol, vortex to mix, then add 1 / 3 volume of supernatant chloroform, vortex to mix, centrifuge at 12,500 rpm for 9 min;

[0044] 4) Take 1100μl of supernatant and add 1 / 2 volume of supernatant to water-saturated phenol, vortex to mix, then add 1 / 2 volume of supernatant to chloroform and vortex to mix, centrifuge at 12500rpm for 9min;

[0045] 5) Take 900μl of supernatant and add equal volume of chloroform, vortex to mix, centrifuge at 12,500 rpm for 10min, take 800μl of supernatant and add equal volume of isop...

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 plant bleeding sap RNA extracting method. The plant bleeding sap RNA extracting method comprises the following steps that freeze frying is conducted on plant bleeding sap, and SDS is added to perform water bathing; water saturated phenol is added and stirred, and then chloroform is added and stirred evenly for centrifugation; supernatant is taken into a novel centrifugation tube and added with water saturated phenol, even stirring is performed, then chloroform is added and stirred evenly for centrifugation; supernatant is taken and added with chloroform, even stirring is performed for centrifugation, and supernatant is taken and added with the same volume of isopropanol for standing; centrifugation is performed, the supernatant is poured off, 80% of ethyl alcohol is adopted for washing and precipitation, 100% of ethyl alcohol is adopted for rewashing, centrifugation and air drying are performed for RNA precipitation, and DEPC water is added. The plant bleeding sap RNA extracting method can inhibit degradation of the RNA in bleeding sap and improve the extracting efficiency of the RNA in bleeding sap. RNA extracting procedures are simplified, the obtained RNA amount is increased, and the amount of RNA extracted from the bleeding sap can meet the inverse transcription demand.

Description

technical field [0001] The invention relates to the technical field of agriculture, in particular to a method for extracting RNA from plant wound fluid. Background technique [0002] At present, the methods for extracting RNA from plant tissues are very mature, but the effect of using these methods to extract RNA from grape, cucumber and other plant wound fluids is not satisfactory, and the following problems arise: 1) RNA cannot be extracted; 2) RNA in wound fluid degrades quickly ; 3) The amount of RNA presented is not sufficient for reverse transcription. Contents of the invention [0003] Purpose of the invention: the purpose of the invention is to provide a method for extracting RNA from plant wound sap. [0004] Technical solution: In order to solve the above technical problems, the present invention provides a method for extracting RNA from plant wound fluid, comprising the following steps: [0005] 1) Freeze-dry the plant wound fluid to 1 / 8-1 / 10 of the original v...

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 解振强王剑贾思振魏跃冯英娜刘叶琼
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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