Cancer Genomics Center

Total RNA sequencing (Total RNA-seq)

Total RNA sequencing service is able to profile both coding and long non-coding transcripts (>100bp). Long non-coding RNAs (lncRNAs) are a moderately abundant fraction of the eukaryotic transcriptome, including lincRNAs (intergentic lncRNAs), intronic, anti-sense, sense and bidirectional lncRNAs. Effects of lncRNAs show evidence on multiple cellular functions and perform as prime targets on the regulation of gene transcription, post-transcriptional modifications, and epigenetics. In total RNA-seq pipeline, mRNAs and lncRNAs were enriched by depleting rRNA from the samples as the figure below.

Sample submission requirements

Service	Sample Type	Recommend Amount	Minium Amount	Minium concentration	Quality
Total RNA-seq	Total RNA	1 ug	200 ng	40 ng/ul	Recommend: RIN>7 Required: RIN>2.3 DV200 >66%

• RIN stands for RNA integrity number, which is evaluated by measuring the ratio of 28S:18S
• DV200 evaluates the percentage of fragments >200 nucleotides
• Total RNA-seq may not be available for some species
• The Core has the rights to determine whether the samples are qualified for the experiment
• Please check our Core (cgc@uth.tmc.edu or 713-500-7933) for more information.

Service Workflow

Useful documents

• NEBNext rRNA Depletion Kit V2
• NEBNext Ultra II Directional RNA Library Prep Kit for Illumina
• DV200 Index for Assessing RNA Integrity in Next-Generation Sequencing