PacBio Iso-Seq full-length non-chimeric transcriptome data of Drosohphila melanogaster ARL0007_XX_tra_lof, 2-4 kb fraction
Iso-Seq libraries from each size fraction were run on a Pacific Biosciences Sequel using Sequel Sequencing Reagent 3.0 and a movie collection time of 20 hours. Iso-Seq library reads were converted to circular consensus sequence (CCS) reads and analyzed with the SMRTLink v8.0 Iso-Seq analysis pipeline to identify full-length non-chimeric CCS reads with single end primer sequences and poly-A tail.
Iso-Seq libraries were created using the Pacific Biosciences Iso-Seq protocol for Sequel Systems and the Clontech SMARTer PCR cDNA Synthesis Kit. Two libraries were prepared from each sample using 600-1000 ng total RNA per library.  The size ranges for the two libraries were approximately 0.8-2 kb and 2-4 kb. Ten cycles of PCR were used for 1000 ng input and 12 cycles for 600 ng input. The amplification cycling conditions were: initial denaturation at 98°C for 30s, followed by 10-12 cycles at 98°C for 10 seconds, 65°C for 15 seconds, 68°C for 10 minutes, and a final extension at 68°C for 5 minutes. SMRTbell libraries were created using the Pacific Biosciences SMRTbell Template Kit.