ITS amplicon sequencing service provides mycobial community profiling. We sequence total DNA extracted from complex matrixes (such as stool, soil, food) to obtain the identification and the abundance of Yeasts and Fungi in the sample.
What we do
- DNA amplification of the ITS2 or ITS1 region with White et al. primers; click here for detailed information on primer sequences.
- Barcoding with Illumina index and multiplexing;
- Sequencing on Illumina Miseq with 300 bp paired-end approach.
- DNA extraction starting from stool, foods (i.e. cheese, yogurt, fermented beverages, etc), soil (20 €/sample). Click here for more information on this service.
- We can provide sterile tubes with preservative buffer for sample shipment at room temperature (5 €/tube).
Output (click on the analysis name to read more about its method and deliverables):
1) Basic Offer: delivery of raw reads in fastq format (about 50,000 reads);
2) Taxonomic Analysis: Basic Offer + reads quality filtering and taxonomic assignments with UNITE database and abundance calculation. An example of the output is available here.
- DNA concentration between 3 and 10 ng/ul (verified by means PCR with primers for fungi/yeasts: see how to check your genomic DNA); it is customer responsibility to send the DNA of interest only;
- DNA volume between 30 and 100 ul provided in 0,2 mL strip-tubes or 96-wells plate with caps (depending on the number of samples);
- In case of 96-wells plate, H11-H12 wells must be empty. Please complete the following file (Template.xls) with samples ID and send it by e-mail to email@example.com.
We test the input DNA with a PCR using universal primers and Illumina-based protocol. Samples not yielding any PCR product will be considered as not-compliant, thus the customer can decide either to stop (paying 10 euros/sample for the PCR done) or to continue with the protocol (but we don’t guarantee any result).
Shipment suggestion: in order to avoid DNA degradation during prolonged storage and shipment we suggest to send the samples in refrigerated packages, at 4°C or -20°C. It is customer responsibility to send the samples by means the most appropriate mode.
Results are usually delivered in about 10 weeks from sample arrival, depending on the working queue and sample compliance to requirements.
Contact us for more info at firstname.lastname@example.org, at +39 049 0995752 or using the following form: