GSM2151697: TP1-KO HSC; Mus musculus; RNA-Seq
For samples 1-8, total RNAs were isolated using RNeasy Plus Mini (for non-HSCs) or Micro (for HSCs) Kit (Qiagen). For samples 9-14, standard ChIP assays was performed. For samples 1-8, Two rounds of polyA+ selection was performed, followed by conversion to cDNAs. Due to the very low amounts, RNAs from the donor-derived LT-HSCs were amplified using Ovation RNA-seq System V2 (Nugen) before library preparation. No amplification was performed for RNAs from non-HSCs. We used the mRNA library generation kits per manufacturer’s instructions (Agilent, Santa Clara, CA). The indexed cDNA libraries were quantitated using qPCR in a Roche LightCycler 480 with the Kapa Biosystems kit for library quantitation (Kapa Biosystems, Woburn, MA) prior to cluster generation. For samples 9-14, input and ChIP DNAs were used to construct libraries for sequencing following Illumina’s standard protocol.
GEO Accession
GSM2151697