Method for rapidly analyzing anaerobic ammonia oxidizing bacteria flora
An anammox bacteria, rapid analysis technology, applied in microorganism-based methods, biochemical equipment and methods, microbial determination/inspection, etc. Quickly analyze problems such as easy digitization, cost reduction, and high repeatability
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
specific Embodiment approach 1
[0024] Embodiment 1: The method for rapid analysis of anammox bacteria in this embodiment is carried out according to the following steps:
[0025] 1. Use the kit to extract the DNA of the sample to be analyzed;
[0026] 2. Specific PCR amplification: take the DNA of the sample as the template, and use Pla46F and 630R as the primers to carry out the first PCR amplification to obtain the amplification product A. The amplification product A is purified by the DNA purification kit to obtain purified product, and then use the purified product as a template and AMX368F and AMX820R as primers to carry out a second PCR amplification to obtain amplification product B;
[0027] 3. Restriction endonuclease digestion: The amplified product B was digested with 1U of mung bean nuclease for 30 min, and the digested amplified product B was purified with a PCR product purification kit, and then MspI and RsaI were used to purify the purified amplified product B is subjected to double-enzymati...
specific Embodiment approach 2
[0033] Specific embodiment 2: This embodiment further describes the first PCR amplification in step 2 of the method for rapid analysis of anammox bacteria described in specific embodiment 1, and the first PCR amplification in step 2 The reaction system was a 50 μL reaction system, consisting of the following components: 10-50 ng of sample DNA, 1 μL primer Pla46F, 1 μL primer 630R, 5 μL 10×Buffer, 4 μL dNTPs, 1 μL Taq enzyme and the balance of ddH 2 O; PCR amplification conditions were as follows: denaturation at 94°C for 5 min, denaturation at 94°C for 30s, annealing at 60°C for 30s, extension at 72°C for 1 min, a total of 30 cycles, then extension at 72°C for 10 min, and incubation at 4°C. Others are the same as the first embodiment.
specific Embodiment approach 3
[0034] Embodiment 3: This embodiment further describes the second PCR amplification in step 2 of the method for rapid analysis of anammox bacteria in Embodiment 1, and the second PCR amplification in step 2 The reaction system is a 50 μL reaction system, which consists of the following components: 10-50 ng of purified product, 1 μL of primer AMX368F, 1 μL of primer AMX820R, 5 μL of 10×Buffer, 4 μL of dNTPs, 1 μL of Taq enzyme and the balance of ddH2 O; PCR amplification conditions were: denaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, extension at 72 °C for 1 min, a total of 30 cycles, and then extended at 72 °C for 10 min, and incubated at 4 °C. Others are the same as the first embodiment.
PUM
Login to view more Abstract
Description
Claims
Application Information
Login to view more - R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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