Fungal Absolute Quantitative Sequencing

Conventional internal transcribed spacer (ITS) amplicon sequencing analysis technology can meet the needs of fungal identification and phylogenetic analysis, but it cannot obtain absolute quantitative information of fungi through this technology. Based on the experience of bacterial absolute quantitative sequencing, Creative BioMart Microbe has established a fungal absolute quantitative sequencing technology platform that combines ITS amplicon sequencing technology and quantitative PCR (qPCR) technology to meet the needs of customers for absolute quantitative analysis of fungi.

Fungal Absolute Quantitative Sequencing Platform

ITS sequence refers to the transcription interval between 18S, 5.8S, 28S rDNA sequences on fungal rDNA. 18S, 5.8S, 28S rDNA sequences are highly conserved in evolution, while ITS sequences mutate rapidly in nature. ITS sequences show sequence polymorphisms in most fungi, with relatively consistent within species and obvious differences between species. Therefore, ITS amplicon sequencing technology is often used for fungal identification and phylogenetic analysis. Similar to the 16S rRNA amplicon sequencing technology, the ITS amplicon sequencing technology can be used to analyze the relative abundance of microbial communities, but this sometimes fails to meet the research needs of microbiologists.

The fungal absolute quantitative sequencing technology platform we established combines ITS amplicon sequencing technology with qPCR technology, which can make up for the deficiencies in qPCR technology, such as high sample quality requirements, difficult primer design, unstable experimental results, and adverse effects of PCR inhibitors, and ultimately obtain accuracy of results High absolute quantitative analysis results.

Creative BioMart Microbe's Bacterial Absolute Quantitative Sequencing Platform Can Help Customers in The Following Aspects:

• Alpha diversity analysis
• Beta diversity analysis
• OTU clustering analysis
• OTU absolute quantification
• Community composition analysis

Sequencing Scheme

• The recommended regions for absolute quantitative fungal sequencing are ITS1, ITS2
• ≥150,000 effective sequence per sample
• Raw data Q30 more than 70%, Clean data 30 more than 80%

Technical Advantages of Absolute Quantitative Sequencing

• High throughput, more than 100000 reads per sample
• Realize the detection of trace fungi in the microbial community
• Simultaneously carry out relative quantitative analysis and absolute quantitative analysis. Realize joint comparative analysis of microbial communities
• Exclude the influence of PCR inhibitors on quantitative results

Sample Requirements

Pre-extracted DNA (in order to ensure the accuracy and reliability of the analysis results, customers need to ensure that the DNA samples provided meet the requirements, or directly perform DNA extraction by our technicians): The total sample volume is required to be > 20 μL, and the number of ITS copies in DNA samples is higher than 2E+03 copies/μL.

Why Choose Creative BioMart Microbe

• Unique quantitative sequencing platform
• Accurate, reliable and efficient fungal quantitative sequencing analysis
• Provide professional and authoritative analysis reports
• One-stop service and reasonable price

Workflow of Fungal Absolute Quantitative Sequencing Service

The fungal quantitative sequencing platform established by Creative BioMart Microbe makes up for the shortcomings of qPCR technology and ITS amplicon sequencing technology, enabling researchers to achieve absolute quantitative analysis of fungi. If you want to know more about our fungal quantitative sequencing technology, please feel free to contact us. You can also choose to conduct service-related consultations through our inquiry form, and our staff will reply to you as soon as possible.