Taking single-cell RNA-seq by STORM: Nanoliter-scale high-resolution total single-cell transcriptome profiling in primary cryopreserved tissue

Miniaturization of genomic library preparation methods has been shown to significantly decrease both costs and labor associated with next-generation sequencing, without significantly sacrificing the quality of data obtained compared to non-miniaturized results. By reducing reaction volumes, the number of samples or single cells that can be prepared per kit is increased, making miniaturization an attractive approach to: 1) reduce the cost per library/cell and 2) take an assay from low throughput to high throughput. Additionally, it is of great importance that a protocol be amenable to preserved tissue sample sources, such as cryopreservation, since acquisition of fresh primary tissues is often unattainable. In an ideal world, the protocol would also be robust to loss of key cell populations (e.g., intermediate cell types) and sufficiently flexible to target populations of interest for transcriptional profiling.

With these aims in mind, research and core facilities at the Van Andel Institute developed STORM-seq (Single-cell TOtal Rna Miniaturized sequencing) as a collaborative effort with Takara Bio and adapted to the SPT Labtech Mosquito HV.

STORM-seq is a nanoliter-scale plate-based single-cell total RNA and ribosomal RNA depletion approach to yield high-resolution single-cell libraries that are comparable to full volume reactions from both cryopreserved human and mouse tissues.

In this webinar, hear how the SPT Labtech Mosquito HV liquid handler was used to miniaturize and increase throughput of the Takara Bio SMART-Seq® Stranded Kit, to yield robust single-cell total RNA libraries from flow-sorted cells, as well as the troubleshooting/optimization challenges and how these were overcome. Finally, the miniaturized single-cell whole-transcriptome data acquired via this method will be highlighted.

Key learning objectives

• How use of the Mosquito HV can help in obtaining single-cell RNA data
• Troubleshooting hurdles of adapting a library prep method to an automated miniaturized platform
• In-depth single-cell profiling of specific tissue

Who should attend?

• Genomics core labs
• Researchers interested in genomics, single-cell profiling, or epigenetics