微生物组/Microbial diversity / amplified multiplon sequencing

Microbial diversity / amplified multiplon sequencing


Technical introduction

The sequencing of microorganism amplification is mainly through direct amplification of specific regions of the total environmental DNA, bypassing the bottleneck of microbial culture, and evaluating the distribution, abundance and community composition of the multi sample environment. With the development of high throughput sequencing technology, the sequencing of microorganism amplification has become one of the main research methods for the comparison and analysis of environmental microbial communities.

According to different research needs, the 16S high variable region is generally selected to distinguish the bacteria and the Archaea community of the environmental samples. The ITS region is amplified to evaluate the fungal community, and the study of the eukaryotic microorganism community, such as the protozoa, can be realized by the sequencing of the 18S high variable region. In addition, specific functional gene sequencing can be used to reveal the distribution of environmental microbial communities with specific functions.

The Owen gene has a standard experimental process, experienced production R & D personnel, professional information analysis team, running international delivery cycle and high quality customer service. Combined with Illumina MiSeq sequencing technology focused on environmental microbial research, it has always provided high quality and efficient sequencing services for microbial community research.

technology roadmap

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Suggestions for sending samples

浓度 (Qubit)体积总量基因组完整性
≥20ng/μL≥15ng/μL≥0.3μgDNA 无降解

technical parameter

测序平台测序策略数据量周期
Illumina MiSeqPE30020,000 raw tags30个自然日

case analysis

Case 1. Distribution of plaque on the upper gums of deciduous teeth and permanent teeth in children during tooth replacement period [1]

As age increases, oral microbes are constantly changing, and tooth replacement is a very meaningful node in oral microbiology research.

In this study, the microbial community analysis process of the eween professional environmental microorganism was used to reveal the distribution of microbial diversity in the teeth and the teeth of the permanent teeth. At the same time, the determination of the core microorganism also provided more research on the growth trend of oral microorganism with age.

In the study, MiSeq sequencing was carried out on the gingival plaque of different dental parts of 20 children's teeth, and the average 18,05 high quality sequence was obtained on the average of each sample. After the analysis of species annotation, it was found that five main categories, such as deformable bacteria and thick wall bacteria, were the most important components of the bacteria group. Further analysis showed that more than half of the low abundance species had significant differences in the abundance between different parts. Over 20% of OTUs was found only in the permanent teeth and the deciduous teeth. The diversity of the community distribution in different parts was mainly reflected in the enrichment of actinomycetes in the permanent teeth and the concentration of the dense spirals in the teeth.



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Fig. 1. comparison of microbial community distribution between deciduous teeth and permanent teeth

Case two, high throughput sequencing technology reveals microbial diversity in rice root system [2]

The mutualism of plant roots and microbes is closely related to plant nutrient utilization, growth and development and disease control. In this study, the microbial community in different regions of rice roots was studied in greenhouse and field, and the network relationship between microorganism and methane cycle and carbon cycle in rice root community was revealed. At the same time, the similarity of the level distribution of the community gate in the rice root microorganism group also hints the study on the terrestrial plant community. The guiding significance of the study.

Through MiSeq sequencing of root, root and Rhizosphere 16SV45 region, it was found that the composition of root microbes in the greenhouse was mainly influenced by soil types and varieties. Under the condition of field, the geographical location, cultivation conditions and varieties affected the diversity of microbial diversity. The study found a large number of microorganisms involved in methane cycle related microorganisms, most of which lack classification information. This study supplements and generalizes the empirical data of the community through the network analysis of related OTUs, and can help identify the associated marker microbes.

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Fig. 2. OTUs abundance network analysis reveals the distribution of methane related microorganisms.

Reference

[1]. Shi W, Qin M, Chen F, et al. Supragingival Microbial Profiles of Permanent and Deciduous Teeth in Children with Mixed Dentition. PloS one, 2016, 11(1): e0146938.
[2]. Edwards J, Johnson C,  et al. Structure, variation, and assembly of the root-associated microbiomes of rice. Proceedings of the National Academy of Sciences, 2015, 112(8): E911-E920.

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