RNA-Seq of Lycium barbarum: green fruit
The total RNA were isolated using the RNAplant Plus Reagent Kit (TIANGEN, Beijing, China) following the manufacturers instructions. The quality, quantity and integrity of the total RNA were assessed by a NanoPhotometer spectrophotometer (IMPLEN, CA, USA), the Qubit RNA Assay Kit in Qubit 2.0 Flurometer (Life Technologies, CA, USA) and the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA), respectively. Briefly, 1.5 g of RNA per sample was used as input material for the RNA sample preparations. Firstly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. First strand cDNA was synthesized using random hexamer primer and M-MLV Reverse Transcriptase (RNase H-). Second strand cDNA synthesis was subsequently performed using DNA polymerase I and RNase H. After cDNA library construction, clusters with satisfactory quality were generated according to the manufacturers recommendation. The library preparations were sequenced on an Illumina HiseqTM 2500 at the Novogene Bioinformatics Institute in Tianjin, China.