Targetamp nano g biotin arna labeling kit

The expression profiles of the genes related to ACh metabolism and signaling transduction pathways were analyzed using the Illumina iScan system with MouseRef-8 Expression BeadChips (Illumina, San Diego, CA, USA) that contain probes that detect over 24 000 transcripts. Total RNAs were isolated by Sepasol-RNA I super solution (Nacalai Tesque, Kyoto, Japan) from the striatum and globus pallidus in wild-type and DD mice. We used the area that includes the neostriatum, ventral striatum, and globus pallidus but not thalamus or hypothalamus in the present study. Total RNA (50 ng) was then applied to RNA amplification that was performed with the TargetAmp Nano-g Biotin-aRNA Labeling Kit (EPICENTRE Biotechnologies, Madison, WI, USA) according to the instructions of the manufacturer. Briefly, first-strand cDNA was synthesized with T7-oligo (dT) primer from total RNAs, and second-strand cDNA synthesis was then performed with synthesized first-strand cDNA. The in vitro transcription reaction was performed with the product of second-strand cDNA synthesis as the template for producing biotin-cRNAs by incorporating biotin-UTP into the RNA transcripts. Finally, 750 ng of biotin-cRNA was hybridized to the MouseRef-8 Expression BeadChips and then reacted with streptavidin-Cy3. The expression intensity of the transcripts on the BeadChips was detected with an Illumina iScan reader.