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<p>We present an optimised protocol for single-nucleus isolation from RNAlater-preserved prostate cancer (PC) resections that enables transcriptomically robust snRNA-seq. Our streamlined procedure employs bead-mill homogenization and high-salt lysis to generate nuclei suspensions, eliminating the need for FACS or density-gradient purification. The protocol yielded high-quality libraries (Q30 ≥ 90%) with minimal ambient RNA. Analysis of 3,365 nuclei identified eight distinct cellular compartments, preserving the full cellular repertoire, including luminal, stromal, and immune lineages. These findings demonstrate that snRNA-seq is feasible in RNAlater-preserved PC specimens enabling routine analysis of clinical samples and expanding the applicability of single-cell genomics in translational research.</p>

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RNAlater-compatible protocol for nuclei isolation from radical prostatectomy prostate cancer resections, enabling single-nucleus resolution RNA-seq

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年6月15日

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  • Magdalena Julita Krystkiewicz-Orzechowska,
  • Dorota Anusewicz,
  • Katarzyna Kośla,
  • Izabela Baryła,
  • Agata Sakowicz,
  • Michał Olczak,
  • Mateusz Mroczek,
  • Marek Lipiński,
  • Andrzej K. Bednarek

摘要

We present an optimised protocol for single-nucleus isolation from RNAlater-preserved prostate cancer (PC) resections that enables transcriptomically robust snRNA-seq. Our streamlined procedure employs bead-mill homogenization and high-salt lysis to generate nuclei suspensions, eliminating the need for FACS or density-gradient purification. The protocol yielded high-quality libraries (Q30 ≥ 90%) with minimal ambient RNA. Analysis of 3,365 nuclei identified eight distinct cellular compartments, preserving the full cellular repertoire, including luminal, stromal, and immune lineages. These findings demonstrate that snRNA-seq is feasible in RNAlater-preserved PC specimens enabling routine analysis of clinical samples and expanding the applicability of single-cell genomics in translational research.