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.
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