PRJNA858999
PacBio IsoSeq data from Pleurodeles waltl female adult brain
PacBio Isoseq data generated from female adult brain tissue from Pleurodeles waltl. Tissue was harvested from the brain of a female adult P. waltl. Tissue was immediately frozen on dry ice and stored at -80C. Approximately 25 mg of brain tissue was then used for tissue pulverization in liquid nitrogen. Tissue powder was used for RNA extraction using Total RNA Purification Kit (Cat. 17200 from Norgen Biotek) as per manufacturer's recommendation. Genomic DNA was removed via on column DNA removal using Norgen RNAse free DNAase kit (Cat. 25710). RNA quality was analyzed via bioanalyzer and had a RIN value of 10. For IsoSeq, cDNA was synthesized using SMARTer PCR cDNA (Clonetech, CA). One IsoSeq library was prepared. Sequencing was performed on a single SMRT cell using PacBio Sequel II. This produced 3,946,27 reads with a mean number of passes of 19, and average read length of 3,618 bp. Data was then fed into IsoSeq3 to generate fasta outputs of high quality and low quality reads. This generated 287,253 polished high-quality isoforms, and 2,802 polished low-quality isoforms. The high quality fasta output was taken for further use as a reference transcriptome.