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scRNA-seq data of human nuclei collected from the temporal cortex of 17 individuals.

Transcriptome analysis of single cells has tremendously increased our understanding of cellular heterogeneity in complex tissues. However, whole-cell transcriptomics cannot be applied to frozen tissue due to poor cell recovery after freezing. Human tissue is usually limited to tissue banks containing frozen material, and since single nuclei can be readily isolated from frozen tissues, human cell nuclei can substitute whole-cell analysis to extract single cell transcriptomes from human cells. Despite the importance of human tissue analysis, single-nucleus RNA-sequencing protocols are less developed than those for whole cells. SMARTSeq, either in form of custom-made SMARTSeq2 or as part of the Fluidigm C1 platform, provides a high resolution view of the transcriptome, has proven effective for whole-cell analysis and has recently been adapted to nuclei. Here, we show that the standard SMARTSeq2 protocol applied to single nuclei biases the amplification of single-nucleus transcriptomes due to unspecific capturing of transcripts, which can comprise the majority of reads in single-nucleus RNA-sequencing libraries. We demonstrate that modification of the SMARTSeq2 protocol with respect to template switching and cDNA amplification conditions generates sequencing libraries of higher complexity, allows for detection of more genes and reduces biases of the amplified transcriptome. In addition, modification of SMARTSeq2 chemistry allows for gene detection at shallower sequencing depth, thus increasing efficiency and decreasing cost.

Click on a Dataset ID in the table below to learn more, and to find out who to contact about access to these data

Dataset ID Description Technology Samples
EGAD00001006575 NextSeq 500 19
Publications Citations
Identification of epilepsy-associated neuronal subtypes and gene expression underlying epileptogenesis.
Nat Commun 11: 2020 5038
77
An overview of human single-cell RNA sequencing studies in neurobiological disease.
Neurobiol Dis 184: 2023 106201
3