CRISPRclean Plus Stranded Total RNA Prep with rRNA Depletion (Human, Mouse, Rat, Pan Bacteria)
Complex samples may have a mixture of bacterial and human cells, all actively expressing ribosomal RNA. CRISPRclean™ Plus depletes over 200 bacteria species and human rRNA sequences to enable identification of lower expressing, biologically relevant transcripts.
CRISPRclean Plus Stranded Total RNA Prep with rRNA Depletion Kit
CRISPR-based ribodepletion strategy optimized with stranded RNA prep increases coverage of lower-expressing transcripts from complex biological samples.
Streamlined CRISPRclean workflow: stranded total RNA library prep with ribodepletion
Ideal for analysis of samples containing complex mixtures of eukaryotic and bacterial rRNA
3 to 7-fold increase in coverage of SARS-CoV-2 and other genomes in nasopharyngeal (NSP) samples
Single workflow to detect viral genomic data, microbiome composition, co-infections, and host gene expression
CRISPRclean Plus Workflow
CRISPRclean Plus Stranded Total RNA Prep with rRNA Depletion (Human, Mouse, Rat, Pan Bacteria) is a streamlined workflow from total RNA to sequencing-ready, strand-specific libraries in 7 steps with multiple safe stopping points.
>80% depletion of eukaryotic and bacterial rRNA in clinical NSP samples
Before and after depletion comparison showed efficient >80% rRNA depletion of eukaryotic and bacterial using clinical NSP samples across different Ct values from negative to 38.93, which has very low viral load.
Top 10 high abundance organisms fold change after depletion
Top ten high abundance organisms in depleted sample S8 (Ct value 26.94). High abundant organisms on average increased 10-fold reads per million compared to undepleted samples.
More gene counts in human gene expression
Gene expression signals before and after depletion are highly correlated with linearity R2 at 0.8589. R2 close to 1.0 shows that human gene expression is conserved between undepleted and depleted sample S7. Note that there are more gene counts for the depleted sample compared to the undepleted sample.
Increased human gene detection sensitivity
More genes were detected after depletion compared to undepleted samples. Read count cutoff of 80 was used as CPM of 1.
Increased bacterial detection sensitivity
More bacterial species are detected when samples are depleted with CRISPRclean.
KIT1016 CRISPRclean Plus Stranded Total RNA Prep with rRNA Depletion (Human, Mouse, Rat, Pan Bacteria) for preparing 24 samples
|Application||Infectious disease surveillance|
|Assay time||9 hours|
|Hands-on time||3.5 hours|
|Sample types||Complex samples with mixtures of eukaryotes and prokaryotes including nasopharyngeal, saliva, gut, skin, and fecal.|
|Input quantity||5ng - 100ng|
|Depletion mechanism||CRISPR-Cas9 mediated|
|Strand specificity||>98% directional and strand specific|
|Compatible species||Human, mouse, rat, 212 bacteria|
|Designed to deplete||Human 5S, 5.8S, 18S and 28S, 45S rRNA precursor, mitochondrial 12S and 16S rRNA genes. 212 bacteria representing all phyla: 5S, 16S, and 23S rRNA gene.|
|Multiplex||Up to 96 unique dual index adapter barcodes|
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For the US patent, the patent number is US 10,604,802 entitled Genome Fractioning. The patent publication is available at https://patentimages.storage.googleapis.com/22/2a/ef/6657ca336c9c08/US10604802.pdf
For the EP patent, the patent number is EP3102722 entitled Genome Fractioning. The patent publication is available at https://patentimages.storage.googleapis.com/a7/54/96/52ed494ab92016/EP3102722B1.pdf