Method for detecting rumen microorganisms of Holstein cows

A technology of rumen microorganisms and microorganisms, which is applied in the field of detection of animal stomach microorganisms, can solve the problems of long sequencing time, complicated operation, and inaccurate measurement results, and achieve the effect of accurate and comprehensive detection results and reduced detection time

Inactive Publication Date: 2015-07-22
XUZHOU NORMAL UNIVERSITY
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Problems solved by technology

The first and second generation sequencing technologies take a long time to sequence, the measurement r

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  • Method for detecting rumen microorganisms of Holstein cows
  • Method for detecting rumen microorganisms of Holstein cows
  • Method for detecting rumen microorganisms of Holstein cows

Examples

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Embodiment 1

[0031] Embodiment 1: the method of the present invention is: extract the rumen fluid of Holstein milk cow, extract the microorganism DNA wherein and determine its microorganism type and relative quantity by sequencing; Prepare according to the following operation steps:

[0032] (1) Genomic DNA extraction: extract the rumen fluid of Holstein dairy cows, then extract and purify the microbial DNA group, and use 1% agarose gel electrophoresis to detect the extracted genomic DNA;

[0033] (2) Polymerase chain reaction: Synthesize specific primers with barcodes according to the designated sequencing region; mix the PCR products of the same sample and detect them by 2% agarose gel electrophoresis, and use the genomic DNA mini kit for gel recovery The PCR product was recovered by cutting the gel of the kit, eluted with trisaminomethane, and detected by 2% agarose electrophoresis; all samples were carried out according to the formal experimental conditions, and each sample was repeated...

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Abstract

The invention provides a method for detecting rumen microorganisms of Holstein cows and belongs to methods for detecting rumen microorganisms of animals. According to the method, rumen fluid of the Holstein cows is extracted, DNA of microorganisms in the rumen fluid is extracted and types and relative quantity of microorganisms of the Holstein cows are determined through sequencing; the method comprises the following steps: (1), genome DNA extraction, (2) PCR amplification, (3) fluorescent quantitation, (4) high-throughput sequencing platform library construction, (5) high-throughput sequencing platform sequencing, (6) biological information analysis and (7) sequencing results. According to the method, the types and relative quantity of rumen microorganisms of the Holstein cows can be rapidly, comprehensively and efficiently detected, so that people can conveniently analyze types and quantity of rumen microorganisms of Holstein cows and study absorption digestion mechanisms for absorbing nourishment of rumen microorganisms. The method has the advantages as follows: detection time is shortened, the detection results are accurate and comprehensive, and types and relative quantity of all microorganisms in the Holstein cows can be accurately detected.

Description

technical field [0001] The invention relates to a method for detecting animal stomach microorganisms, in particular to a method for detecting rumen microorganisms of Holstein cows. Background technique [0002] High-throughput sequencing technology (High-throughput sequencing), also known as "next-generation" sequencing technology ("Next-generation" sequencing technology), can perform sequence determination and general reading on hundreds of thousands to several million DNA molecules in parallel at a time. Longer, shorter, etc. are signs. [0003] According to the development history, influence, sequencing principle and technology, there are mainly the following types: massively parallel signature sequencing (Massively Parallel Signature Sequencing, MPSS), polymerase cloning (Polony Sequencing), 454 pyrosequencing (454pyrosequencing), Illumina (Solexa) sequencing, ABI SOLiD sequencing, Ion semiconductor sequencing, DNA nanoball sequencing, etc. The first and second generat...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06
CPCC12Q1/689C12Q1/686C12Q2561/101
Inventor 王秀颖朱春婷温洪宇
Owner XUZHOU NORMAL UNIVERSITY
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