PacBio sequencing of rice: whole plant 20 days after germination
The Poly(A) RNA was isolated from the total RNA using the oligo d(T) magnetic bead binding method and the Poly(A) PuristTM Kit (Thermo Scientific, http://www.lifetechnologies.com/uk/en/home/brands/ambion.html). 0.5 ?g of poly (A) RNA was reversely transcribed into cDNA using the SMART cDNA Library Construction Kit, which uses Long-Distance PCR method to generate full-length double strand cDNA, after first-strand cDNA synthesis (Takara Biotechnology). The SMRT bell library inserted with cDNAs of 1-2 kb, collected through 1.0% agarose gel electrophoresis, was constructed using 3.0 ?g size-selected cDNA with the Pacific Biosciences DNA Template Prep Kit 2.0 (Pacific Biosciences).