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Sequencing | 16S/18S/ITS Sequencing

Microbial Profiling – 16S/18S/ITS Amplicon Sequencing

16S/18S/ITS amplicon sequencing is an established method for microbial identification and phylogeny studies of environmental or microbiome samples. 16S rRNA sequencing is used for bacterial and arechea identification, whereas 18S rRNA and ITS (Internal Transcribed Spacer) amplicon sequencing is used for the identification of fungi. 

Microbial Analysis Services

We offer microbial analysis services using pre-designed primers to specific regions or we can sequence custom regions using a tailed amplicon approach.  For our routine microbial analysis services we have the following primer pairs:

  • 16S_V1-V3 (27f-519r)
  • 16S_V4 (515f-806r)
  • 16S_V3-V4 (341f-805r)
  • 18S_V4 (V4_18SNextF-V4_18SNextR)
  • 18S_V9 (1391f-EukBr)
  • ITS (ITS1F-ITS4)
  • ITS2 (fITS7-ITS4)
  • A16S_V1-V3 (A2f-519r)

The average price per sample is less than $60 AUS when submitting a batch of 94 samples with significant price breaks for larger submissions. Please contact us for a quote. Further details on our primer selection can be found below.

For detailed instructions on how to submit samples for this service please read our Sample Submission Guide. In brief we require:

  • 15-20ul of DNA at a concentration of 5-10ng/ul (based on Qubit/PicoGreen quantification). 
  • DNA of high purity (260/280, 260/230 >~1.8).  
  • Samples should be submitted in 96-well PCR plates in consecutive columns with no gaps between samples.
  •  94 samples is the maximum number that can be submitted per plate. Two wells are required for positive and negative controls.
  • Samples must be amplifiable by PCR. Please check this prior to submission and provide a gel photo. 
  • If your samples require any special conditions or additives (i.e. BSA, DMSO) to amplify, please let us know at the time of submission.

Notes on Primer Selection

16S-V4 and 18S-V9 are used by the Earth Microbiome Project for their 16S (original) & 18S protocols.

16S_V1-V3, ITS, 18S_V9, and A16S_V1-V3 are used in the Bioplatforms Australia Biome of Australia Soil Environments (BASE) project.

16S_V1-V3, 18S_V4, and A16S_V1-V3 are used in the Bioplatforms Australia Marine Microbes project.

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