| Popis: |
Deep sequencing of cDNA prepared from total RNA (RNA-Seq) or mRNA (mRNA-Seq) has become the method of choice for transcript profiling, discovery of novel transcripts, and identification of alternative splicing events. However, standard whole-transcriptome approaches to RNA-Seq face a significant challenge, as the vast majority of reads map to rRNA. One solution—poly(A) enrichment—does not capture several biologically relevant RNA species, such as microRNA and also requires large amounts of Total RNA. Whole blood RNA samples pose an even bigger challenge due to the presence of reticulocyte globin transcripts that can constitute up to 76% of total mRNA and, unless removed along with the ribosomal RNA transcripts, the detection of low-abundance mRNA transcripts is further compromised. To overcome these challenges, Epicentre developed Globin-Zero™ technology for blood derived RNA. The Globin Zero Gold kit removes rRNA and globin mRNA from intact and fragmented blood total RNA samples (human, mouse and rat), and an improved, more user-friendly version of the ScriptSeq™ RNA-Seq library preparation method. Directional cDNA libraries with ∼98% strandedness can be prepared in about 4 hours, in a single-tube workflow, from either intact or fragmented Globin-Zero treated RNA samples, without the need for end-polishing, adaptor ligation, cDNA fragmentation or gel size-selection. The combined Globin-Zero and ScriptSeq workflow generates cluster-ready NGS libraries in ∼6 hours. Globin-Zero-treated samples contain. |
