100mg of spleen tissue from each of the 150 individuals was disrupted using Lysing Matrix D tubes (MP Biomedicals, Irvine, CA) on a FastPrep homogenizer (MP Biomedicals) and resuspended in RLT buffer (Qiagen, Hilden, Germany) for total RNA preparation. RNA was purified using an RNeasy Mini kit and treated with an RNase-free DNase Set. Concentration and purity was measured using A260/280 and an Agilent 2100 Bioanalyzer. 1ug of total RNA was used for each sample.
Summarization: PLIER, Background: PM-GCBG, Normalization: Global Median
Total RNA was extracted using TRIzol (Invitrogen) and further purified and DNase I treated using an RNeasy Mini kit (Qiagen) and RNase-Free DNase Set (Quiagen), according to the manufacturer protocols. RNA concentration and purity was determined through measurement of A260/280 ratios with a NanoDrop ND-1000 Spectrophotometer. Confirmation of RNA quality was assessed using the Agilent 2100 Bioanalyzer. RNA samples were immediately frozen and stored at -80C. 1ug of total RNA was used for each sample.
A backcross population was established between experimental autoimmune encephalomyelitis (EAE)-susceptible DA and EAE-resistant PVG rats. Rats were subsequently immunized with a 200ul inoculum of 25-30ug of recombinant myelin oligodendrocyte glycoprotein in PBS emulsified in Incomplete Freunds Adjuvant subcutaneously at the tail base to induce an autoimmune response. Thirty-five days later, spleens were removed and snap frozen in liquid nitrogen and stored in -70oC until use.