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Cancer Genomics Center

Oncogenomics

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.


total-rna-sequencing-ultra-low-input6 total rna-sequencing-diagram



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

service-workflow-total-rna-sequencing-ultra-low-input-total-rna-sequencing-diagram



Useful documents