RNA integrity. RNA integrity number (RIN) was measured with the Agilent RNA 6000 Pico kit using the Agilent 2100 Bioanalyser. Chips were prepared for all human brain RNA according to the manufacturer’s instructions. The RNA samples and ladder were prepared and kept on ice until use. XXX was measured recorded in Table 2.5. RIN number less than ~ 5 was excluded from data analysis because it is below the acceptable range for good RNA quality. Briefly, 550 μl of the RNA 6000 Pico gel matrix was placed on a spin filter, and centrifuged at 1, 500 g for 10 min at room temperature. This was then divided into 65 μl aliquots. After the addition of 1 μl of the RNA 6000 Pico dye concentrate to the 65 ul aliquot of filtered gel, the gel-dye mix was vortexed and centrifuged at 13, 000 g for 10 min. A RNA ladder consisting of a mixture of RNA with known concentration was run after the addition of 5 μl marker to the ladder and sample xxxxx. 1 μl (3 ng/μl) RNA of each RNA sample was added in the 11 designated sample xxxxx, followed by the addition of 1 μl of diluted RNA 6000 ladder in the ladder well. The kit consists of interconnected microchannels that separate RNA fragments (18S and 28S for eukaryotic RNA) based on their size when driven electrophoretically. The eukaryotic 18S and 28S RNA are detected and plotted on an electropherogram graph. To determine the RIN number, the area under the RNA electropherogram is calculated and converted to concentration values using the ladder’s concentration/area ratio.
