title: RNA-Seq of Argyranthemum frutescens
design: Total RNA was used as input material for the RNA sample preparations. Briefly, mRNA with polyA tail was enriched from total RNA using Oligo(dT) attached magnetic beads . Fragmentation was carried out using divalent cations under elevated temperature in Fragmentation Buffer . First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase , then use RNaseH to degrade the RNA. Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and dNTPs. Remaining overhangs were converted into blunt ends via exonuclease / polymerase activities. After adenylation of 3 ends of DNA fragments. Adaptor with hairpin loop structure were ligated to prepare for hybridization. To select cDNA fragments of preferentially 370~426 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter , Beverly , USA). Then PCR was performed with Phusion High-Fidelity DNA polymerase , Universal PCR primers and Index (X) Primer.
library_name: AF7
identifiers.namespace: SUB15154152
identifiers.id: AF7