CRISPRclean Stranded Total RNA Prep with rRNA Depletion

Specific, unbiased rRNA depletion and library prep for RNA-Seq

Product Description

RNA samples frequently contain up to 90% ribosomal RNA. Transcriptome sequencing without removing these overabundant rRNA sequences reduces sequencing capacity and makes it harder to detect lower expressing, biologically relevant transcripts. See how CRISPRclean can help your research with effective rRNA removal.

• Effective single tube, multi-species depletion with CRISPR-Cas9 mediated technology
• Increases sensitivity to detect lower expressing, biologically relevant transcripts
• Optimized library prep and depletion workflow generates high quality representation of transcripts
• Consistent full-length, uniform transcript coverage, and high library complexity
• For Human, Mouse, and Rat

How it works

The workflow begins with RNA fragmentation through high-temperature incubation and continues to first and second strand synthesis, converting RNA fragments into cDNA libraries. The assay achieves 98% strand specificity by incorporating dUTP during second strand synthesis. Adenylation modifies the 3’ ends of the double-stranded cDNA and dATP prepares the library for adapter ligation. After ligating the unique dual index adapters, the library is ready for depletion. CRISPR depletion happens in two successive incubations. The first cleaves bacterial rRNAs and the second cleaves eukaryotic rRNAs. Cas9 and guide RNAs are combined to form the ribonucleoprotein complex that cuts the targeted rRNA sequences. Cleaved rRNA cannot be amplified and is removed through size selection with magnetic beads.

Specifications

Related Product: CRISPRclean Unique Dual Index Adapter Plate for RNA Prep (Set A)

Specific and unbiased

CRISPRclean Stranded Total RNA Prep with rRNA Depletion produces extremely low library bias. ERCC reads counts were highly correlated, between depleted (y-axis) and undepleted libraries (x-axis), indicating the CRISPRclean rRNA depletion method is highly specific, unbiased, and accurate. This allows for gene expression measurements to be more accurately represented than those that would be obtained from an undepleted sample. Illumina RiboZero Plus Depletion paired with Jumpcode’s stranded RNA prep shows lower ERCC read count correlation and higher bias.

CRISPRclean Depletion                                                                            

RiboZero Plus Depletion

Fig.1. CRISPRclean versus RiboZero Plus Depletion

Performance Data

Fig.2. >90% rRNA depletion across mammalian species. Universal reference RNAs (Human, mouse, rat, chicken, cow, and dog) were used to assess performance of the ribodepletion across multiple species.