title: GSM2065904: PacBio_Spombe; Schizosaccharomyces pombe; RNA-Seq
library_construction_protocol: Nascent RNA was prepared using a nascent RNA isolation protocol for yeast (Carrillo Oesterreich et al. 2010) and Phenol:Chloroform:IAA, 25:24:1, pH 6.6 extraction. Nascent RNA was 3’ end ligated to the DNA adaptor (/5rApp/NNNNNCTGTAGGCACCATCAAT/3ddC/) and reverse transcribed into cDNA with a slight modification of the SMARTer PCR cDNA Synthesis Kit (Clontech). A custom primer reverse-complement to the 3’ end adaptor was used (AAGCAGTGGTATCAACGCAGAGTACATTGATGGTGCCTACAG). Maximal 1 µg of RNA was used per reaction. The subsequent low cycle PCR was carried out with the Advantage 2 PCR Kit according to the instructions provided by the kit. Double-stranded cDNA was further used for Pacific Biosciences library preparation and sequencing with standard protocols and kit from Pacific Biosciences (SMRTbell™ Template Prep Kit 1.0). SMRT DNA sequencing (Pacific Biosciences)
attributes.tag: GEO Accession
attributes.value: GSM2065904