A pilot single-nucleus RNA sequencing experiment was undertaken to compare single-cell versus single-nuclear results from a matched sample. The biopsy from a 58-year-old male was collected fresh and divided into 8 segments, evenly distributed to be processed fresh for single cell RNA sequencing as above, and the remainder was flash frozen in liquid nitrogen. The sample was later retrieved from liquid nitrogen and processed on dry ice according to the protocol in88 (link) with a lysis buffer containing: 0.32 mM sucrose (BioShop SUC507.1), 5 mM CaCl2 (VWR, 97062-820), 3 mM MgCl2 (Thermo Fisher AM9530G), 20 mM Tris-HCl pH 7.5 (Thermo Fisher, 15567027), 0.1% TritonX-100 (Sigma Aldrich T8787-50ML), 0.1 mM EDTA pH 8.0 (Thermo Fisher AM9260G), 40 U/ml Protector RNAse inhibitor (Sigma Aldrich 3335399001) in UltraPure DNAse/RNAse-free water (Thermo Fisher 10977015). The nuclei were captured and sequenced using 10X Genomics Single Cell 3’ v3 Reagents as above.
Suc507
Manufactured by Avantor
The SUC507.1 is a laboratory equipment designed for general scientific applications. It serves as a core component for various experiments and research activities. The product specifications and technical details are available upon request.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using suc507
1
Single-Nucleus RNA Sequencing of Frozen Tissue
2
Single-Nucleus RNA Sequencing Pilot
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A pilot single-nucleus RNA sequencing experiment was undertaken to compare single cell versus single nuclear results from a matched sample. The biopsy was collected fresh and divided into 8 segments, evenly distributed to be processed fresh for single cell RNA sequencing as above, and the remainder was flash frozen in liquid nitrogen. The sample was later retrieved from liquid nitrogen and processed on dry ice according to the protocol in 74 with a lysis buffer containing: 0.32 mM sucrose (BioShop SUC507.1), 5 mM CaCl2 (VWR, 97062-820), 3 mM MgCl2 (Thermo Fisher AM9530G), 20 mM Tris-HCl pH 7.5 (Thermo Fisher, 15567027), 0.1% TritonX-100 (Sigma Aldrich T8787-50ML), 0.1 mM EDTA pH 8.0 (Thermo Fisher AM9260G), 40 U/ml Protector RNAse inhibitor (Sigma Aldrich 3335399001) in UltraPure DNAse/RNAse-free water (Thermo Fisher 10977015). The nuclei were captured and sequenced using 10X Genomics Single Cell 3' v3
Reagents as above.
Reagents as above.
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