Advanced Results from Sample to Data
DNA microarrays have traditionally dominated applications involving transcriptome analysis, profiling of protein-DNA interactions, and small-scale and large scale genetic variation. However, the processing speed, knowledge requirement for sequences being interrogated, problematic cross-hybridization, and the analog nature of the microarray signal has made it difficult to detect and quantify low-abundance RNA species. These low abundance species include non-polyA+ tailed mRNA, preprocessed RNA, potentially regulatory RNA, and other RNA transcripts of unknown function in assessing the true whole transcriptome. The transcriptome is defined as the complete collection of transcribed elements of the genome and contains mRNA transcripts and non-mRNA transcripts. Since large rRNA constitutes 90–95% RNA species in total RNA, whole transcriptome analysis without any contamination from rRNA is very difficult using existing RNA isolation methods including microarray.
The Ribominus™ Eukaryote Kit for RNA-Seq was developed to provide a superior method for true whole transcriptome isolation through the selective depletion of ribosomal RNA (rRNA). The unique and patented Locked Nucleic Acid (LNA) probe based RiboMinus™ system selectively depletes up to 99.9% of rRNA, including the 5S, 5.8S, 18S and 28S rRNA components in order to increase specificity across multiple organisms (Figure 2, 3).
Unbiased depletion of rRNA is achieved with no effect from the differing expression levels of multiple genes (figure 4).The remaining transcriptome RNA can then be used for digital interrogation of the transcriptome using next generation sequencing technology (ex. ABI SOLiD, Illumina Solexa). Compared to traditional polyA selection methods RiboMinus™ depleted samples provide superior depth and breadth of coverage across long genes thereby increasing the amount and accuracy of the sequencing information obtained (figure 5). This same effect has been demonstrated across multiple genes (figure 6). The RiboMinus™ method is not dependent on the polyadenylation status or presence of a 5′-cap structure on the RNA as with existing methods that only offer a partial isolation of the transcriptome.
Unbiased depletion of rRNA is achieved with no effect from the differing expression levels of multiple genes (figure 4).The remaining transcriptome RNA can then be used for digital interrogation of the transcriptome using next generation sequencing technology (ex. ABI SOLiD, Illumina Solexa). Compared to traditional polyA selection methods RiboMinus™ depleted samples provide superior depth and breadth of coverage across long genes thereby increasing the amount and accuracy of the sequencing information obtained (figure 5). This same effect has been demonstrated across multiple genes (figure 6). The RiboMinus™ method is not dependent on the polyadenylation status or presence of a 5′-cap structure on the RNA as with existing methods that only offer a partial isolation of the transcriptome.
For Research Use Only. Not for use in diagnostic procedures.