Truseq pe cluster kit version 4 cbot hs

Fruit skin from the self-rooted and grafted cucumbers (12 d after blooming) were used for RNA extraction. Total RNA was extracted using the RNeasy Plant Mini Kit (Qiagen), assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, Santa Clara, CA, USA), and the RNA was sent to Beijing Novogene Bioinformatics Technology Company (Beijing, China) for sequencing and bioinformatics analysis. The Illumina NEBNext UltraTM RNA Library Prep Kit (Illumina, San Diego, CA, USA) was used for library construction. To select cDNA fragments 370–420 bp in length, the library fragments were purified using the AMPure XP system (Beckman Coulter, Beverly, MA, USA). PCR was performed using the Phusion High-Fidelity DNA polymerase, Universal PCR primers, and an Index (X) Primer. After PCR amplification and purification of the PCR products, the library quality was assessed using an Agilent Bioanalyzer 2100. After cluster generation using the TruSeq PE Cluster Kit version 4-cBot-HS (Illumina, San Diego, CA, USA), the library preparations were sequenced on an Illumina Novaseq platform, and 150 bp paired-end reads were generated.

The Plant Quick RNA Isolation Kit (Huayueyang Biotech, Beijing, China) was used to extract total RNA from tea leaves. The concentration and quality of RNA were determined using Agilent 2,100 Bioanalyzer (California, CA, United States) and Nano-Drop 2000 ultra-micro spectrophotometer (ThermoFisher, Waltham, MA, United States). Then, 1 μg total RNA was used to prepare the RNA-seq transcriptome library according to the manual of NEBNext Ultra™ RNA Library Prep Kit for Illumina (NEB, Massachusetts, MA, United States). To select cDNA fragments of preferentially 240 bp in length, the library fragments were purified using the AMPure XP system (Beckman Coulter, Beverly, United States). After PCR amplification and purification of the PCR products, the library quality was assessed using Agilent Bioanalyzer 2,100. After cluster generation using TruSeq PE Cluster Kit version 4-cBot-HS (Illumina, California, CA, United States), the library was sequenced on the Illumina 2,500 sequencer for paired-end sequencing to obtain raw reads.