Illumina HiSeq 2000 paired end sequencing; Effects of human RelA transgene on murine macrophage inflammatory responses
Effects of human RelA transgene on murine macrophage inflammatory responses
Bone marrow was isolated from mouse femurs and aliquots prepared in 90% v/v foetal bovine serum (FBS) containing 10% v/v dimethyl sulfoxide (DMSO). Vials were stored in liquid nitrogen until further use. Bone marrow cells were cultured in vitro in the presence of 50 ng/mL recombinant macrophage colony-stimulating factor (MCSF) (Peprotech; London, UK) to induce differentiation of monocytes to macrophages Isolation and purification of RNA were performed using the RNeasy mini kit (Qiagen), as per manufacturer instructions. Strand-specific sequencing libraries were prepared with the TruSeq stranded Total RNA kit (Illumina) from 1 Âµg total RNA of each sample