GSM4944277: Rubinstein-Taybi syndrome type 1 rep 1; Mus musculus; RNA-Seq
Peripheral blood was obtained from facial vein bleed and CD19+ cells were isolated by positive selection using magnetic microbeads (Miltenyi 130-052-201) per manufacturer protocols and placed on ice. 100k-500k cells were spun at 300-500g for 5 min at 4', homogenized in Trizol (Invitrogen 15596018) and stored at -80 until extraction. RNA was phase separated using standard protocols followed by purification using the Direct-zol RNA microprep kit (Zymo R2060) with an on-column DNAse step per manufacturer directions. RNA was then quantified using Quant-iT RiboGreen RNA Assay Kit (ThermoFisher R11490) or Qubit RNA HS Assay Kit (ThermoFisher Q32852), and quality checked by Bioanalyzer with RNA 6000 Pico Kit (Agilent 5067-1513). mRNA was isolated using (NEBNext Poly(A) mRNA Magnetic Isolation Module; NEB #E7490). Libraries were generated using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB E7760/E7765) per manufacturer protocols. Library size and quality were determined using BioAnalyzer with DNA High Sensitivity reagents (Agilent 5067-4626), and concentration was determined using Qubit dsDNA HS Assay Kit (ThermoFisher Q32851) and KAPA Library Quantification Kit for qPCR (KAPA KK4824).
GEO Accession
GSM4944277