Cancer Genomics Center

Ultra-low input mRNA sequencing (Ultra-low input mRNA-seq)

This sequencing service is able to generate high-quality libraries directly from live cells (100-1,000 cells) or 1-10 ng of total RNA. It provides the excellent sensitivity needed to obtain full-length transcript information from high-quality total RNA (RIN >8) or intact cells by using oligo(dT) priming. Through even gene-body coverage and accurate representation of GC-rich transcripts, this chemistry enables reliable analysis of transcript isoforms, gene fusions, point mutations, and more.

Highlights

• provide a solution for the samples with limit material
• an alternative method to supplement single-cell RNA sequencing with lower price

Sample submission requirements

Service	Sample Type	Recommended Amount	Minimum Amount	Minimum Concentration	Quality
Ultra-low input mRNA Seq	Total RNA	10 ng	7 ng	1 ng/ul	Recommend: RIN>8 Required: RIN>7
	Live Cells	1000-100 cells	100 cells		Cell viability: >85%

• RIN stands for RNA integrity number, which is evaluated by measuring the ratio of 28S:18S
• Concentration must be measured by Qubit
• We recommend to sort live cells at UTH flow cytometry core https://www.uth.edu/imm/service-centers/flow-cytometry/
• The core has the rights to determine if the samples are qualified for the experiment
• Please check the core (cgc@uth.tmc.edu or 713-500-7933) for the detail information.

Service Workflow

Useful documents

• SMART-Seq v4 Ultra Low Input RNA KIt
• SMART-Seq v4 PLUS Kit