Single-nucleus suspensions are easily generated from fresh and frozen tissue using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator with Heaters. Our solution combines a gentle mechanical disruption of the tissues and efficient cell lysis to release intact nuclei in minutes and from up to eight samples in parallel. The extraction is performed at low temperatures; assuring high-quality nuclei for reliable single-nucleus RNA-seq.
No matter the tissue type, high numbers of nuclei can be extracted using the Nuclei Extraction Buffer and the gentleMACS Octo Dissociator with Heaters. We have validated a wide range of fresh and frozen tissues including snap frozen mouse brain, liver, heart and kidney, syngeneic mouse tumors, and OCT embedded human tumors.
Our technology enables the generation of intact and high-quality nuclei suspensions through gentle dissociation and mild cell lysis for reliable downstream nuclei analysis.
The figure shows the numbers of extracted nuclei per mg of dissociated mouse brain (1) and mouse kidney (2), as indicated in the figure legend. Single-nucleus suspensions were obtained from fresh and frozen tissues by using the Nuclei Extraction Buffer and the gentleMACS Octo Dissociator with Heaters. Nuclei were stained with DAPI and analyzed by flow cytometry using the MACSQuant® 10 Analyzer.
Single-nuclei sequencing applications require high quality single-nuclei suspensions. Performed in a cold environment, our method allows the obtention of intact nuclei with highly preserved RNA for optimal subsequent genomic applications.
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