P02RY19095583
Populus ussuriensis single-molecule long read isoform sequencing
Total RNA from 9 biological replicates was pooled in equal amounts and 1 g of the pooled RNA was used for cDNA synthesis and SMRT bell library construction.The purified RNA was reverse transcribed into cDNA using the SMARTer PCR cDNA Synthesis Kit (Clontech Laboratories, Inc. Mountain View, CA, USA). cDNA was amplified using the Kapa HiFi PCR kit (Kapa Biosystems, Wilmington, MA, USA). Size selection was carried out on a BluePippin (Sage Science, Beverly, MA, USA) and 1-2 kb, 2-3 kb, 3-6 kb, and 5-10 kb fractions were collected. After size selection, the collected cDNA fractions were treated with DNA damage repair mix, followed by end repair and ligation of SMRT adapters using the PacBio SMRTbell Template Prep Kit (Pacific Biosciences, Menlo Park, CA, USA) to construct PacBio libraries.
P02RY19095583
SUB7848877
P02RY19095583