title: GSM6435645: CK2,Plant tissue; Juglans regia; RNA-Seq
library_construction_protocol: Total RNA was extracted by phenol-chloroform method, followed by removal of genomic DNA with DNase, RNase inhibitor and other reagents. 1.3 ug of total RNA was used to construct sequencing libraries. The fragmented mRNA was used as a template and random oligonucleotides were used as primers to synthesize the first strand of cDNA in M-MuLV reverse transcriptase system, followed by degradation of RNA strands with RNaseH and synthesis of the second strand of cDNA with dNTPs under DNA polymerase I system. The purified double-stranded cDNA was end-repaired, A-tailed and ligated with sequencing connectors. cDNAs of about 250-300bp were screened with AMPure XP beads, PCR amplified and PCR products were purified again using AMPure XP beads to finally obtain libraries. The kit used for library construction is NEBNext® Ultra™ RNA Library Prep Kit for Illumina®.