GSM9359507: CAGE_L_00146.BC04,Rottweiler,Cerebral_cortex_pool; Canis lupus familiaris; RNA-Seq
CAGE libraries were prepared using 1 μg of total RNA from each sample.The CAGE experiment requires at least 10 μl of sample to proceed. Therefore, tissue pools were created for samples with insufficient volume. We begin with at least 10 μl of volume for each sample. CAGE libraries were prepared following the previously described protocol by Takahashi et al. (Takahashi et al., 2012). In each library, we used eight barcodes (CTT, GAT, ACT, ATG, AGT, TGG, GTA, and GCC) for eight RNA samples. Single-strand cDNA was barcoded by adding a 5' linker containing a 5' barcode sequence and the EcoP15I sequence. A 3′ linker with Illumina primer sequence was added to the barcoded cDNA fragment. The size of the resulting CAGE libraries ranged between 97 and 103 bp. Libraries were single-read sequenced at 50 cycles using the HiSeq 2500 Illumina System. The HiSeq SR Rapid Cluster Kit v2 (Illumina) and the HiSeq Rapid Duo cBot Sample Loading Kit (Illumina) were used to sequence CAGE libraries in the rapid mode.