Revvity Sites Globally

Select your location.

*e-commerce not available for this region.

Australia

Austria

Belgium

Brazil *

Canada

China *

Denmark

Finland

France

Germany

Hong Kong (China) *

India *

Ireland

Italy

Japan *

Luxembourg

Mexico *

Netherlands

Norway

Philippines *

Republic of Korea *

Singapore *

Spain

Sweden

Switzerland

Thailand *

United Kingdom

United States

NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human)

The NEXTFLEX™ Cas9-gRNA Globin Depletion Enzyme (Human) is a standalone, flexible CRISPR-Cas9 module designed to remove globin-derived sequences from human blood RNA-seq libraries. It can be dropped into existing bulk or single-cell RNA-seq workflows at the double-stranded DNA stage, reducing hemoglobin background, so a greater share of reads is directed toward biologically informative transcripts and improved transcriptome coverage.

Supplied as a pre-complexed Cas9-guide RNA mixture, the NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) contains 30 guide RNAs targeting the HBA1, HBA2, HBB, and HBD loci. A unit is defined as the amount required to deplete 1 - 10 ng of target dsDNA. Developed to be highly cost-competitive and widely compatible, it delivers efficient globin depletion without locking you into a particular library prep kit.

View product information

Feature	Specification
Automation Compatible	Yes
Product Group	RNA Depletion

View product information

Product variants

Unit Size: 50 μL

Part #:

NOVA-802100

Unit Size: 250 μL

Part #:

NOVA-802500

For research use only. Not for use in diagnostic procedures.

Request more information

Product information

Overview

Globin transcripts can account for a large fraction of reads in whole blood RNA sequencing libraries, which can mask low-abundance, biologically relevant transcripts. The NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) is a cost-effective, workflow-independent, and highly specific enzyme that reduces globin-derived reads, so more sequencing capacity is directed toward informative genes, without requiring you to move to a new library prep kit.

It uses a DASH-style (Depletion of Abundant Sequences by Hybridization) CRISPR-Cas9 depletion approach, with a precomplexed Cas9 and pool of guides that cleave human globin genes at any dsDNA stage. Whether you use your own prep method or a commercial kit, this module gives you clean, reproducible depletion of globin sequences and improved coverage of non-globin transcripts.

Cas9-gRNA-mediated targeted removal of human globin genes
Ready-to-use: provided as a Cas9-gRNA complex with integrated RNase inhibitor and 10X Cas9 Buffer
Fast: short one-hour reaction time with only 5 minutes of hands-on time
Convenient: easily fits into any library prep workflow
Flexible: compatible with short- and long-read sequencing
Automation-compatible: easily incorporated into manual or automated workflows

Additional product information

Cas9-gRNA-driven targeted globin depletion

This enzyme leverages Jumpcode® DepleteX™ technology to selectively remove globin-derived sequences at the double-stranded DNA (dsDNA) stage, giving you the flexibility to deplete either immediately after second-strand synthesis or on completed libraries, depending on your workflow. The pre-complexed Cas9–guide RNA pool contains 30 unique guides against the human HBA1, HBA2, HBB, and HBD loci, selectively cleaving hemoglobin-derived fragments from blood RNA-seq libraries while preserving non-globin transcripts. This targeted approach reduces globin carryover and increases the efficiency of blood transcriptome sequencing without altering your upstream chemistry.

A unit of NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) is defined as the amount needed to deplete 1–10 ng of target dsDNA, assuming a library size of ≥ 400 bp (typically corresponding to an insert size of ~300 bp or greater).

Stacked bar chart comparing RNA-seq libraries before and after NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) treatment.

Figure 1. Globin depletion with NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) increases informative reads in blood RNA-seq libraries. Stacked bar chart shows the percentage of mapped reads assigned to globin genes (purple) and non-globin genes (green) in untreated and Cas9-depleted libraries. In untreated libraries, globin transcripts account for nearly half of mapped reads, whereas after Cas9 globin depletion almost all mapped reads align to non-globin genes.

Compatible with virtually any RNA-seq workflow

Because it is supplied as a standalone Cas9–gRNA complex rather than built into a fixed library prep kit, the NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) fits into almost any RNA-seq strategy. If your samples have reached the double-stranded DNA stage, you can apply globin depletion. Operating at the dsDNA level avoids constraints around RNA input quality or chemistry and allows the module to integrate cleanly with existing blood RNA-seq workflows, regardless of which reagents or sequencing platform you use, including long- and short-read workflows.

Need an end-to-end RNA-seq solution?

This globin depletion module is fully compatible with NEXTFLEX Rapid Directional RNA-seq kits . Whether you are working with high-quality RNA or more degraded samples, NEXTFLEX RNA-seq workflows deliver robust performance, high mapping rates, and flexible input requirements. Pair this module with a complete NEXTFLEX prep kit to streamline your workflow, increase the fraction of informative reads, and receive end-to-end support including globin removal and sequencing-ready libraries.

Specifications

Other specifications

Automation Compatible	Yes
Product Group	RNA Depletion
Shipping Conditions	Shipped in Dry Ice
Unit Size	50 μL

FAQs

What are the benefits of using CRISPR-based globin depletion in RNA-seq libraries?

CRISPR-based depletion uses Cas9–guide RNA complexes to cut unwanted globin-derived fragments at the double-stranded DNA (dsDNA) stage of the library. This helps redirect sequencing depth from highly expressed hemoglobin transcripts to lower-abundance, biologically informative genes, while preserving overall transcript diversity, including fragments derived from challenging or partially degraded samples.

Why deplete globin sequences at the dsDNA library stage instead of at the RNA stage? Most globin depletion strategies act directly on RNA, which can be problematic for low-input or degraded samples because RNA is inherently labile. By removing globin-derived fragments after they have been converted to dsDNA, the NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) minimizes the risk of losing rare or fragmented transcripts during depletion and is easier to drop into existing workflows without additional RNA-handling steps.
Which species and sample types are supported for globin depletion?

The NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) is designed for human globin genes and targets HBA1, HBA2, HBB, and HBD. It is intended for libraries prepared from human whole blood or other blood-derived RNA where globin transcripts represent a substantial fraction of reads. It is not validated for non-human species.
What library prep kits and sequencing platforms are compatible with DASH?

Because depletion occurs at the dsDNA stage, this module is compatible with a wide range of bulk and single-cell RNA-seq workflows for short- or long-read sequencing, including NEXTFLEX Rapid Directional RNA-seq kit 2.0 and other commonly used commercial or in-house protocols. As long as your library has progressed to dsDNA with fragment sizes of roughly 300 bp inserts (≥400 bp library size), the enzyme can typically be integrated with a simple incubation and bead-based cleanup.
Is the NEXTFLEX Cas9-gRNA Globin Depletion Enzyme suitable for degraded or low-input samples?

Yes. Because the depletion is performed on dsDNA libraries rather than on RNA, the approach is well suited to libraries generated from partially degraded or low-input RNA, where additional manipulation at the RNA stage could result in loss of material. Performance will still depend on the quality and complexity of the starting library, so standard QC is recommended.
How much globin reduction can I expect?

In internal testing on human whole blood RNA-seq libraries, untreated samples showed approximately half of mapped reads originating from globin genes, whereas libraries treated with the NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) had >99% of mapped reads aligning to non-globin genes (see Figure 1). Actual performance may vary depending on sample type, library preparation method, and sequencing depth.
Does globin depletion affect non-globin transcripts or introduce bias?

The guide set is programmed to target the major human globin loci (HBA1, HBA2, HBB, HBD), focusing Cas9 cleavage on hemoglobin-derived fragments so that non-globin genes are retained. As with any targeted depletion strategy, we recommend evaluating coverage metrics and gene-detection performance in your specific workflow, especially when working with unusual library designs or extreme GC content.
Can this module be used in DASH-style workflows or alongside other Cas9-based depletion modules?

Yes. DASH refers to CRISPR-Cas9 depletion of abundant sequences in sequencing libraries. The NEXTFLEX Cas9-gRNA Globin Depletion Enzyme (Human) is a ready-to-use Cas9–gRNA complex for DASH-style removal of globin sequences from human libraries with a one-hour incubation at the dsDNA stage. In principle it can be combined with other Cas9-based depletion modules in the same general workflow, though reaction conditions, order of steps, and cleanup procedures may need to be optimized for your specific application.