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

Method for specific separation of Acidobacteria from soil

An acid-bacterial, specific technology, applied in the intersection of modern molecular biology technology and microbial technology, to achieve strong specificity, reduce false amplification, and reduce competition for nutrients

Inactive Publication Date: 2017-08-18
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there is still a lack of high-efficiency primers with sufficient specificity to complete the specific amplification of Acidobacterium 16S rDNA.

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 specific separation of Acidobacteria from soil
  • Method for specific separation of Acidobacteria from soil
  • Method for specific separation of Acidobacteria from soil

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Collect forest soil from Mount Wuyi in Jiangxi Province, my country, pass through a 2mm sieve, and store in a refrigerator at 4°C. Weigh 5g of the soil sample and place it in a sterilized Erlenmeyer flask filled with glass beads, add 45mL of physiological saline, and shake at 150rpm for 2h. Then the suspension was allowed to stand still, the supernatant was taken, and the soil suspension was diluted with a 10-fold volume gradient with physiological saline. Take 0.1ml respectively to dilute to 10 -3 、10 -4 、10 -5 、10 -6 The soil suspension was spread on the solid basic medium plate after adding the improved one, and cultured upside down at 30°C for 3-6 days. Pick the white colony ( figure 1 ), streaked repeatedly on the improved solid basic medium plate until a single colony was isolated.

[0042] The configuration method of the improved liquid basal medium is: KH 2 PO 4 1.5g, MgSO 4 7H 2 0.2g, sucrose 30g, yeast extract 50mg, agar powder 20g (add when config...

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 which combines the modern molecular biological techniques with the traditional microbiology cultivation techniques and carries out specific separation on a basic culture medium plate improved by combining specific primer-PCR based on a gate level to obtain Acidobacteria difficultly cultured in the traditional meaning. The method comprises the following steps: diluting and applying sample suspension liquid onto the improved basic culture medium plate, selecting white single colonies, purifying by continuously using a continuous streak plate method, performing amplification culture on the purified single colonies through a liquid culture medium, extracting genomic DNA, amplifying 16S rDNA fragments by employing the Acidobacteria specific primer PCR, and if a positive amplification product of which the fragment size is 1462 is obtained, showing that Acidobacteria in the sample is screened. According to the invention, gate specific primer amplification is combined with an acid screening plate method, so that cultivable Acidobacteria can be separated from samples effectively.

Description

technical field [0001] The field that the present invention relates to is the intersection field of modern molecular biology technology and microbe technology, specifically a kind of method for specifically isolating acidobacteria based on specific primer-PCR combined with improved basic medium plate. Background technique [0002] Acidobacteria (Acidobacteria) was first discovered by Kishimoto in Japan in 1991. As an important part of the soil microbial community, Acidobacteria is second only to Proteobacteria in relative abundance, accounting for about the total amount of soil bacteria. In some specific environments, such as chestnut rhizosphere soil, as the absolute dominant microbial group in the soil, its relative abundance can reach 65%. Acidobacteria are widely distributed in the environment, including natural environments and some extreme environments and polluted environments, and Acidobacteria play an important ecological role in driving soil material cycle and ecol...

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): C12N1/02C12Q1/68C12Q1/04C12R1/01
CPCC12N1/02C12Q1/689
Inventor 李慧姚飞杨山王志瑞姜勇
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
PatSnap Eureka logo

PatSnap Eureka turns technology decisions into work you can execute. Powered by our Innovation Knowledge Graph, it runs expert workflows across engineering, life sciences, materials and intellectual property. Get your review-ready output in minutes.

X (Twitter)LinkedInYouTubeSubstack

© 2026 PatSnap. All rights reserved. 

Terms of Service

 | 

Privacy policy

 | 

Modern Slavery Act Transparency Statement