Rna rlt buffer

AMA-non-treated or treated central neuron systems of wild-type Drosophila larvae as well as neuron cells of E17 Wistar rat embryos were collected and homogenized in 350 μl RNA RLT buffer (Qiagen)/0.01% 2-mercaptoethanol (Wako, Tokyo, Japan), respectively. Total RNA was purified with RNeasy mini kit (Qiagen). To eliminate genomic DNA contamination, on-column DNA digestion was carried out for each sample with DNase I (Qiagen). Prepared RNA samples were subjected to a HiSEq based RNA-seq by TAKARA (700 million-bp reads). Gene expression of each sample were calculated using IGV Tools (IGV version 2.3.55, Broad Institute, Cambridge, MA, USA), and differential expression of the genes were analyzed with R (version 3.1.3, R Foundation for Statistical Computing, Vienna, Austria). Gene expression was calculated as reads per kilobase of exon per million mapped reads (RPKMs).

SCA1-iPSCs and normal iPSCs were collected and homogenized in 350 µL RNA RLT buffer (Qiagen)/0.01% 2-mercaptoethanol (Wako, Tokyo, Japan). Total RNA was purified with RNeasy mini kit (Qiagen). To eliminate genomic DNA contamination, on-column DNA digestion was conducted for each sample with DNase I (Qiagen). Prepared RNA samples were subjected to a HiSeq-based RNA-seq by TAKARA (700 million bp reads).