1000, 10k and 100k purified mouse kidney FFPE mouse nuclei were split into different 0.5 ml Qubit tube (Invitrogen, Q32856)s with duplicate after epitope retrieval and all of them were washed once with FACT-seq Antibody buffer (20 mM HEPES(K+) pH 7.6 (HEPES: Sigma-Aldrich, H3375; KOH: Sigma-Aldrich, 484016), 150 mM NaCl (Invitrogen, AM9759), 2 mM EDTA (INVITROGEN, AM9260G), 0.5 mM Spermidine (Sigma-Aldrich, S2626), 0.05% Digitonin (Millipore, 300410), 0.01% IGEPAL® CA-630 (Sigma-Aldrich, 13021-50), 1× protease inhibitors (Sigma-Aldrich, 11873580001), 1% BSA (Miltenyi Biotech MACS, 130-091-376)). The H3K27ac FACT-seq sequencing libraries were prepared with same methods in the section of FACT-seq for mouse FFPE nuclei.
Qubit tube
Manufactured by Thermo Fisher Scientific
The Qubit tube is a laboratory equipment designed for the quantification of DNA, RNA, and protein samples. It provides a precise and accurate measurement of the concentration of these biomolecules in a sample.
Automatically generated - may contain errors
2 protocols using qubit tube
1
Preparing FFPE Mouse Kidney Nuclei
2
Nuclei Extraction and H3K27ac FACT-seq Library Prep
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Aliquots of nuclei with epitope retrieval from human GBM or human CRC samples (100 000 cells/0.5 ml Qubit tube (Invitrogen, Q32856)) were washed once with FACT-seq Antibody buffer (20 mM HEPES(K+) pH 7.6 (HEPES: Sigma-Aldrich, H3375; KOH: Sigma-Aldrich, 484016), 150 mM NaCl (Invitrogen, AM9759), 2 mM EDTA (Invitrogen, AM9260G), 0.5 mM spermidine (Sigma-Aldrich, S2626), 0.05% digitonin (Millipore, 300410), 0.01% IGEPAL® CA-630 (Sigma-Aldrich, 13021-50), 1× protease inhibitors (Sigma-Aldrich, 11873580001), 1% BSA (Miltenyi Biotech MACS, 130-091-376)). H3K27ac FACT-seq sequencing libraries were prepared with same methods as in the mouse samples.
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