I.DOT for scalable single-cell RNA sequencing

Author: Tarun Mahto – Application Scientist

High throughput sequencing methods have transformed the field of biology during the last decade. The ability to investigate complete transcriptomes in depth using RNA sequencing (RNA-seq) has fuelled various significant discoveries and has now become a standard method in biomedical research.

RNA-seq, is often performed in bulk, and the results represent an average of gene expression patterns across hundreds to millions of cells; this may mask biologically meaningful changes across cells.

Sequencing the RNA of single cells is one method for addressing this issue. ScRNA-seq enables assessment of fundamental biological features of cell populations and biological systems at unprecedented precision by isolating single cells, collecting their transcripts, and producing sequencing libraries in which the transcripts are mapped to individual cells.

Although several single-cell RNA sequencing (scRNA-seq) approaches have been studied, and their various strengths and shortcomings have been identified researchers must still choose between methods with high cell throughput but low transcript coverage (droplet or combinatorial indexing methods) and methods with high sensitivity and full-length transcript coverage (plate-based methods).

I.DOT, an accurate nanolitre liquid handler, has proven to be a suitable platform for the Smart-seq3xpress technique. It miniaturizes and streamlines the Smart-seq3 process, to minimize reagent input and boost cellular throughput without compromising data quality.

Using the I.DOT, the authors of the paper were able to reduce the reaction volumes of the lysis, reverse transcription (RT), and pre-amplification PCR stages to 1:2, 1:5, and 1:10 of the established quantities and evaluate these conditions on K562 and HEK293FT cells. (See Figure 01-03). As a result, it dramatically decreases costs, saves valuable time, and minimizes plastic consumables.

 

Access the free research article for more information.

Download: https://doi.org/10.1038/s41587-022-01311-4

Read our blog on Sustainable High Throughput Screening in Drug Discovery.

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