Year: 2020
Type: preprint
Abstract: This protocol describes a nuclei isolation method for frozen tissues based on the previously published protocol by Gao et al. and modified by Drokhlyansky et al. It can be used on both healthy and disease tissues and is compatible with droplet-based single-nucleus RNA-Seq technologySlyper et al. This method is part of a toolbox for processing frozen tissue samples forsingle-nucleus RNA-Seq, including the NST (this protocol), CST and TST protocols (all available inprotocols.io). We recommend users to test all three protocols on their tissue type of interest and perform side-by-side comparison of the data generated. A description of the complete toolbox andguidance for testing and selecting methods from the toolbox for processing other tumors can be found in Slyper et al. For the Human Tumor Atlas Pilot Project (HTAPP), this protocol was tested on metastatic breast cancer, ovarian cancer and pediatric neuroblastoma frozen samples. For these cancer types, however, TST was the protocol of choice. When tested on HTAPP samples, NST, in general, underperformed compared to TST and CST based on several QC parameters.
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W4238591670