GSM5665645: CK+B-1; Glycyrrhiza uralensis; RNA-Seq
Glycyrrhiza uralensis Fisch. seedling were collected and ground. Then RNA was extracted and its purity, concentration and integrity were tested. 1ug total RNA was prepared for cDNA libraries using protocol provided by Oxford Nanopore Technologies (ONT). Briefly, SuperScript IV First-Strand Synthesis System (Invitrogen) was used for full length mRNA reverse transcription and following cDNA PCR for 14 circles with LongAmp Tag (NEB). The PCR products were then subjected to FFPE DNA repair and end-repair (NEB) steps and following adaptor ligation using T4 DNA ligase (NEB). Agencourt XP beads was used for DNA purification according to ONT protocol. The final cDNA libraries were added to FLO-MIN109 flowcells and were run on PromethION platform at Biomarker Technology Company (Beijing, China). RNA libraries were prepared for sequencing using standard Illumina protocols