16S Identification

16S Identification

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Technology

The genomic analysis of multiple organisms obtained from a single sample, commonly referred to as ‘metagenomics’, allows insight into the genetic composition of microbial communities including the species that cannot be cultured and isolated in the laboratory.

Third generation sequencing technology, also known as Nanopore Sequencing, has been a revolutionary approach to sequence genomic molecules entirely. Whereas other sequencing technologies allow to obtain just hundreds of base pairs, Nanopore sequencing can cover thousands of them in single reads. This creates an opportunity to sequence a complete PCR product obtained from one or several species within a sample, such as the 16s rRNA gene that has been a standard for taxonomic classification.

Applications

The applications for this service are several, like: the confirmation of previously sequenced samples, discovery of new species, classification of a consortium, studies on environmental samples, studies of changes in the composition of microbiomes over time and more.

Advantages

No need to purify cultures, unless you want the de novo assembly of the full length 16S rRNA.

No need to look at specific variable regions, you can have them all sequenced at the same time.

Abundance species table provided.

Raw read accuracy >Q15 with high sequencing depth

Workflow

We amplify the entire 16S rRNA gene using universal primers 27F and 1492R generating a product ofapproximately 1,500 bp from extracted gDNA.

Later on, this gets sequenced with Nanopore Sequencing, producing full length sequencing data. This amplicon containing variable regions 1 to 9 allows the analysis of this gene across the present prokaryotic organisms in your samples, speeding up the identification and making it cost-effective.

The taxonomic classification is done with Kraken2 and the reference database from NCBI.

Library kit

From Macrogen Europe we recommend to send at least 10μl for optimal results.

Sequencers

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