title: GSM6928341: miRNA-Seq FZX_Le_C3; Litchi chinensis; miRNA-Seq
library_construction_protocol: Total RNA was extracted from the treated leaves by Plant RNA Kit (Promega, LS1040). The small RNA library (sRNA library) was constructed according to the manufacturer's instruction of TrueSeq small RNA library prep kit (Illumina San Diego CA, USA), and 2.5 ng RNA per sample was used as the initial amount. The T4 RNA Ligase 1 was ligated to the 3′ end of the RNA, followed by the ligation of T4 RNA Ligase 2 (truncated) to the 5′ adapter. After that, the RNA was reverse transcribed to synthesize cDNA. Finally, small RNA libraries were obtained by screening these recovered fragments using gel separation technology. The qualities of the library was further tested following these steps: 1) Initial quantification was performed using Qubit 2.0, and the insert size of the library was tested using Agilent 2100. 2) The Q-PCR method was used to accurately quantify the effective concentration of the library (effective library concentration > 2 nM). After the library's quality tests were passed, the library performed pooling according to the amount of target data, followed by sequencing on the Illumina Nova platform.