Cancer Genomics Center

Ultra-low input total RNA sequencing (Ultra-low input total RNA-seq)

This sequencing service is able to provide a solution for those samples which can’t increase the amount and improve the quality, such as FFPE samples. SMART® (Switching Mechanism At 5' end of RNA Template) cDNA synthesis technology (Chenchik et al. 1998) and generates Illumina-compatible libraries via PCR amplification, avoiding the need for adapter ligation. The directionality of the template-switching reaction preserves the strand orientation of the original RNA, making it possible to obtain strand-specific sequencing data from the synthesized cDNA.

Sample Submission requirements

Service	Sample Type	Recommend Amount	Minium Amount	Minimum Concentration	Quality
Ultra-low input total RNA-seq	Total RNA	10 ng	5 ng	1 ng/ul	Must be treated by RNase-free DNase 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
• This service may not be available for some species
• Concentration must be measured by Qubit
• 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 more information

Services Workflow

Useful documents

• SMARTer Stranded Total RNA-seq Kit v2
• Technological note