High Input rRNA depletion Kit
High Input riboPOOLs
Ribodepletion kit for total RNA input of more than 5 µg. Removes unwanted ribosomal RNA from total RNA samples, enabling clearer insights for applications like Direct RNA Sequencing or detecting rare or lowly expressed transcripts.
High Input
Process up to 30µg of total RNA per sample instead of the usual maximum of 1-5µg.
Shorter Workflow
Consolidate what previously required 10 separate reactions into a single step.
Cost-saving
For depleting large amounts of RNA, this kit will be the cheapest option per-reaction.
When Do You Need High-Input rRNA Depletion?
The High-Input riboPOOL rRNA Depletion Kit (5 µg–30 µg input) is developed for RNA sequencing workflows that require large amounts of depleted RNA.
Typical applications include:
- Direct RNA sequencing
- Long-read sequencing
- Detection of rare or lowly expressed RNAs
After depletion, the RNA is well suited for advanced long-read transcriptomics workflows, including Oxford Nanopore Technologies and Pacific Biosciences (PacBio) platforms. As a result, transcript structures and splice variants can be characterized with greater precision.
Because ribosomal RNA is efficiently removed, sequencing capacity is directed toward biologically relevant transcripts. Consequently, rare and low-abundance RNAs are detected more reliably, and overall data quality improves.
In particular, high RNA input is essential for direct RNA sequencing. Since this approach avoids PCR amplification, more starting material is required to generate sufficient signal and sequencing depth. Moreover, this enables applications such as RNA modification analysis (epitranscriptomics).
How Much RNA Is Considered “High Input” for rRNA Depletion?
The riboPOOL High Input Kit expands our proven rRNA depletion technology to process up to 30 μg of total RNA per reaction. This represents a 10-fold increase compared to standard protocols.
As a result, high-input workflows become more cost-efficient while maintaining the same reliable depletion performance. At the same time, the kit remains fully compatible with short- and long-read sequencing platforms. In addition, it supports the broad species coverage that defines the riboPOOL platform.
Validated rRNA removal
Figure 1. Percentage of rRNA reads in total RNA from A549 cells before and after rRNA depletion with the human High Input riboPOOL,
based on Oxford Nanopore Direct RNA Sequencing.
Figure 2. Bioanalyzer data after ribodepletion of 7, 14 and 28 μg of depleted total input RNA of different lengths.
Figure 3. Bioanalyzer profile of human rRNA-depleted RNA, including or excluding the small nucleotide fraction (<200 nucleotides). Column-based purification provides the option to include or exclude the small nucleotide fraction, depending on your needs.
Available riboPOOLs
Single-species riboPOOLs
Specifically designed to deplete rRNAs in single species
Acetabularia ryukyuensi
ID: 620
Aedes albopictus
ID: 547
Amphimedon queenslandica
ID: 511
Anaeramoeba flamelloides
ID: 595
Anopheles gambiae
ID: 597
Apis mellifera
ID: 573
Arabidopsis thaliana
ID: 508
Argiope bruennichi
ID: 574
Azolla filiculoides
ID: 579
Bacillus subtilis
ID: 512
Bemisia tabaci
ID: 580
C. elegans
ID: 539
C. crescentus
ID: 513
Chinchilla lanigera
ID: 514
Chlamydomonas reinhardtii
ID: 571
Clostridium perfringens
ID: 515
Crassostrea gigas
ID: 563
Cryptococcus neoformans
ID: 584
Cupriavidus necator
ID: 602
Danio rerio
ID: 510
Deroceras laeve
ID: 635
Drosophila melanogaster
ID: 507
Emiliania huxleyi
ID: 533
Escherichia coli
ID: 504
Escherichia coli tRNA
ID: 634
Escherichia coli SsrA
ID: 607
Eubacterium limosum
ID: 570
Gallus gallus domesticus
ID: 560
Haloferax volcanii
ID: 516
Human
ID: 554
Human Blood
ID: 594
Human Cytoplasm
ID: 598
Human Globin mRNA
ID: 525
Human tRNA
ID: 617
Ixodes scapularis
ID: 530
Leptinotarsa decemlineata
ID: 572
Mouse tRNA
ID: 622
Nematostella vectensis
ID: 587
Olavius algarvensis
ID: 603
Oryza sativa
ID: 509
Pichia pastoris (fungi)
ID: 519
Pinus sylvestris
ID: 608
Plautia stali
ID: 520
Pseudomonas aeruginosa
ID: 518
Saccharomyces cerevisiae
ID: 505
Salmonella enterica
ID: 522
Schistosoma mansoni
ID: 590
S. pombe
ID: 559
Schmidtea mediterranea
ID: 528
Spodoptera exigua
ID: 585
Sus scrofa Globin mRNA
ID: 636
Staphylococccus aureus
ID: 521
Stenotro-phomonas sp.
ID: 523
Strongyloides ratti
ID: 606
Thalassiosira pseudonana
ID: 596
Ustilago maydis
ID: 524
Varroa destructor
ID: 581
Wolbachia pipientis
ID: 586
Pan riboPOOLs
rRNA depletion for multiple species
Pan-Actinobacteria
ID: 535
Pan-Archaea
ID: 527
Pan-Bacteria
ID: 526
Pan-Bird
ID: 532
Pan-Chlamydia
ID: 611
Pan-Dictyostelia
ID: 609
Pan-Fungi
ID: 543
Pan-Mammal
ID: 541
Pan-Mussel
ID: 589
Pan-Plant
ID: 531
Pan-Prokaryote
ID: 503
Pan-Sponge
ID: 544
Blood parasite
ID: 551
Filamentous fungi
ID: 506
Human-Holobiont
ID: 633
H/M/R
ID: 553
L. orbiculatus & L. aequizonata
ID: 517
Mouse/Rat
ID: 555
Seawater
ID: 568
Willaertia-Naegleria
ID: 610
Combination riboPOOLs
Combine up to 4 riboPOOLs for different species
Custom riboPOOLs
Order a riboPOOL customized to your needs
FAQ: Ribodepletion for more than 5 µg total RNA samples
How much total RNA can I depleted in one reaction?
Up to 30 µg of total RNA.
Can I deviate from the Input amount?
We strongly recommend using a total RNA input between 5 µg and 30 µg for this kit. If you need a kit for a lower input amount, we recommend using our standard riboPOOls.
There is currently no riboPOOL kit available for my species. Is a custom design possible?
Yes. We offer custom design for both standard and high-input riboPOOLs.
Is the High-Input riboPOOL kit compatible with long-read sequencing platforms?
Yes. The kit is compatible with both long and short-read sequencing platform.
What should I use for the cleanup step?
For the high input riboPOOL, the recommended purification strategy is to use a column-based product. We recommend the RNA Clean and Concentrator kit (from Zymo Research).
Is the new kit compatible with degraded RNA or ribosome profiling?
The kit is currently only available for all our standard RNA-Seq riboPOOLs, which are compatible with RNA samples with RIN value of 7 or more.
How much RNA is left after rRNA depletion?
The amount depends on type of sample and depletion efficiency. We regularly recover > 400 ng rRNA-depleted RNA when starting from 30 µg
There is currently no riboPOOL kit available for my species. Is a custom design possible?
Yes. We offer custom design for both standard and high-input riboPOOLs.