Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for extracting aeromonas veronii total RNA

A technology of Aeromonas viridis and an extraction method, which is applied in the field of genetic engineering molecule extraction, can solve the problems of low total RNA concentration and poor effect, and achieves the effect of increasing the concentration

Pending Publication Date: 2019-12-10
HAINAN UNIVERSITY
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the specific steps of extracting the total RNA of Aeromonas verdeii, the most commonly used method is to use the operating procedure issued by the Cold Spring Harbor Laboratory in the United States. However, the classic method of Cold Spring Harbor is used to directly extract the total RNA of Aeromonas verderichi The concentration is extremely low, the effect is not good, and it is often difficult to use for subsequent real-time fluorescent quantitative PCR experiments

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
  • Method for extracting aeromonas veronii total RNA
  • Method for extracting aeromonas veronii total RNA
  • Method for extracting aeromonas veronii total RNA

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0041] The operation of formulation preparation is as follows:

[0042] Before starting, wash all the empty bottles used to store the solution and rinse them once with deionized water, fill them with the prepared reagents, and tighten the bottle caps after autoclaving.

[0043] 1. The preparation process of ethanol (100%) is as follows:

[0044] Note: Pre-cool at -40°C before use.

[0045] Sterilization method: Sterilization is not required.

[0046] 2. The preparation process of PCA(25:24:1) is as follows:

[0047] Preparation method: V (water-saturated phenol): V (chloroform): V (isoamyl alcohol) = 25:24:1;

[0048] Note: It is ready-to-use and prepared in advance in proportion to each use in RNase free 1.5ml EP tubes. For each sample, one tube of 600 μl PCA and one tube of 400 μl PCA need to be prepared. Due to possible stratification, they need to be prepared separately, otherwise the absorption ratio is wrong. After configuration, store temporarily in the dark at 4°C...

no. 2 example

[0098] The method in the first embodiment was used to extract the total RNA of Aeromonas verkirea, and the total RNA was purified using the column type RNA rapid concentration purification kit of B518688 produced by Shanghai Sangong, and then agarose gelation was carried out. Gel electrophoresis verification, the electrophoresis picture is as follows figure 2 indicated, and the concentration of RNA was determined.

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 relates to a method for extracting aeromonas veronii total RNA, and belongs to the technical field of genetic engineering molecule extraction. Aeromonas veronii total RNA extraction is performed through a method provided by an American Cold Spring Harbor Laboratory operation flow, and lysozyme of which the concentration is 5mg / ml + / -5% is added in an extraction step. Compared with aconventional extraction method, the extraction method provided by the invention can enable the concentration of the obtained RNA to be notably increased, and requirements of a subsequent reverse transcription experiment system are met; and the RNA is purified through a column type RNA quick concentration and purification reagent kit of which the type is B518688 and which is produced by Shanghai sangon, the situation that the pollution of gDNA ( genome DNA) is eliminated is indicated, and the problem of RNA degrading can also be solved.

Description

technical field [0001] The invention relates to the technical field of genetic engineering molecule extraction, in particular to an improved method for extracting total RNA from Aeromonas verkirea by adding lysozyme. Background technique [0002] Aeromonas veronii (Aeromonas veronii), a Gram-negative bacteria, rod-shaped cells, facultative anaerobic, respiratory and fermentation two metabolism. [0003] After the total RNA extraction of Aeromonas verkisii, a real-time fluorescent quantitative PCR experiment can be performed. In the specific steps of extracting the total RNA of Aeromonas verdeii, the most commonly used method is to use the operating procedure issued by the Cold Spring Harbor Laboratory in the United States. However, the classic method of Cold Spring Harbor is used to directly extract the total RNA of Aeromonas verderichi The concentration is extremely low, the effect is not good, and it is often difficult to use for subsequent real-time fluorescent quantitat...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12R1/01
CPCC12N15/1003C12Q2521/531C12Q2523/32
Inventor 马香徐一轲刘柱李宏唐燕琼胡康林敏
Owner HAINAN UNIVERSITY