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- W4387306065 abstract "Single-cell and single-nuclei approaches are being extensively used to decipher the cellular heterogeneity of samples and to understand the genetic factors leading to various diseases. While single-cell RNA sequencing is quite straightforward, single-nuclei sequencing requires careful consideration in the isolation & processing of intact nuclei after dissociation. Numerous protocols have been developed for this purpose, but the size and morphology of cardiomyocytes make the process quite complicated.1-3 Previously described processes also included steps involving manual homogenization using a Dounce homogenizer and Fluorescence-activated Cell Sorting.2,4 These steps have an increased propensity to vary from user to user and also produce stress on the cells, which further alters the transcriptional profile of these nuclei. To this end, we devised a simple direct protocol for processing cardiac tissue into intact nuclei for downstream single-nuclei RNA sequencing using the gentleMACS Octo Dissociator. This protocol has been extensively tested with Engineered Heart Tissues (EHTs) and also works well with frozen or fresh cardiac tissues from human biopsies." @default.
- W4387306065 created "2023-10-04" @default.
- W4387306065 creator A5042873961 @default.
- W4387306065 date "2023-07-26" @default.
- W4387306065 modified "2023-10-05" @default.
- W4387306065 title "GONE in 360 seconds: gentleMACS Octo Dissociator-based Nuclei Extraction v1" @default.
- W4387306065 doi "https://doi.org/10.17504/protocols.io.dm6gp3bo8vzp/v1" @default.
- W4387306065 hasPublicationYear "2023" @default.
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