RNA-seq of Cucurbita pepo:Fruit_2_DAP
The extracted RNA was checked using a NanoDrop 2000 (Thermo, CA, USA), and the RNA concentration and integrity were assessed using the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent, CA, USA) to ensure that the RNA Integrity Number (RIN) values were above 7.0. Oligo (dT) beads were used to isolate poly(A) + mRNA, which was fragmented to 250 bp. Fragmentation of the RNA and reverse transcription of double-strand cDNA (ds cDNA) by N6 random primer. The synthesized cDNA was subjected to end-repair and then was 3 adenylated. Adaptors were ligated to the ends of these 3 adenylated cDNA fragments; The ligation products were purified and many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer; Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and DNA ligase; Each cDNA library was sequenced in a single lane of the BGISEQ-500 system with paired-end sequencing length of 100 bp according to the manufacturers instructions at the Beijing Genomics Institute (BGI-Shenzhen, Denmark).
Fruit-set-1