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Nuclei Isolation from Fresh Frozen Brain Tumors for Single-Nucleus RNA-seq and ATAC-seq

Article DOI: 10.3791/61542-v • 06:22 min • August 25th, 2020
August 25th, 2020 •
1, 2, 1, 1,3, 4, 2, 1
1Neurology Clinic and National Center for Tumor Diseases, University Hospital Heidelberg, 2Bioinformatics and Omics Data Analytics, German Cancer Research Center (DKFZ), 3Clinical Cooperation Unit Neurooncology, German Cancer Research Center (DKFZ), 4Division of Experimental Neurosurgery, Department of Neurosurgery, University of Heidelberg, INF 400, Heidelberg, Germany

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Intra-tumoral heterogeneity is an inherent feature of tumors, including gliomas. We developed a simple and efficient protocol that utilizes a combination of buffers and gradient centrifugation to isolate single nuclei from fresh frozen glioma tissues for single nucleus RNA and ATAC sequencing studies.

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Nuclei Isolation Fresh Frozen Brain Tumors Single-nucleus RNA-seq ATAC-seq Transcriptional Studies Epigenetic Studies Tumor Evolution Therapy Effects High-quality Nuclei Stress Reduction Archival Frozen Tumors Ashwin Narayanan Tissue Sample Transfer Tissue Mincing/chopping Nuclei Lysis Buffer Douncing Procedure
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