General information on Ribo Zero Plus Microbiome

What is Ribo-Zero Plus Microbiome?

• This product contains additional rRNA depletion probes, designed to target species found in the microbiome of human adult and infant stool samples.

• Ribo-Zero Plus Microbiome can either be purchased as a standalone product, or bundled with library prep reagents in the Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Microbiome kit, listed on the product page here.

Which other sample types are compatible with Ribo-Zero Plus Microbiome?

• Illumina has reports of effective depletion from skin, vaginal, and oral samples in addition to stool samples, though results may vary.

What probes are included in Ribo-Zero Plus Microbiome?

• The Microbiome probe set includes a total of 1092 probes, including ATCC probes (380), and the existing E. coli and B. subtilis probes (57).

How much input is needed? What quality does it need to be?

• The Ribo-Zero Plus Microbiome protocol is optimized for 25-500 ng of total RNA input.

• RNA is recommended to have a RIN of at least 5.

How to use the Ribo-Zero Plus Microbiome reagents?

• The full protocol is in the updated Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus, Ribo-Zero Plus Microbiome Reference Guide (1000000124514 v03 or later).

• Briefly, DPM (Microbiome Depletion Probe Pool) is added to DP1 (Ribo-Zero Plus Depletion Probe Pool) to deplete all rRNA species in a single step.

What are the sequencing recommendations for Ribo-Zero Plus Microbiome?

• Illumina recommends a 2 x 150 bp read length, rather than 2 x 75 bp, for better analysis results.

How to analyze Ribo-Zero Plus Microbiome sequencing data?

• One option it to use the Microbiome Metatranscriptomics App In BaseSpace Sequence Hub.

  • This app is optimized for data prepared using the Illumina Stranded Total RNA with Ligation, Ribo-Zero Plus Microbiome.

See also:

• Ribo-Zero Plus Microbiome Depletion Kit data sheet.
• Ribosomal RNA depletion for metatranscriptomic studies app note.

For any feedback or questions regarding this article (Illumina Knowledge Article #6974), contact Illumina Technical Support [e-mail].