GSM5566188: Integument of 32DAP in MT rep1; Citrus reticulata; RNA-Seq
RNA was extracted from cell mass captured by laser capture microdissection following the manufacturer's suggested protocol (Arcturus Picopure kit, Kit0204). About 10-20ng total RNA of 36 samples (contained three biological replications) was used for cDNA synthesis and amplification using an Ovation RNA-seq System V2 Kit (Nugen Technologies). The first strand cDNA were synthesized using random hexamer primers and dNTPs, and the second strand cDNA were synthesized using the first strand cDNA as a template with same reaction mix as above. The purification steps were carried out in accordance with the protocol. The product was used to SPIA cDNA amplification following by the purification again with QIAGEN MinElute Reaction Cleanup Kit to guarantee no primer and dNTP remains. The cDNA libraries were sequenced with the paired-end (PE, 2 x 150bp) strategy on the Hiseq X ten platform / NovaSeq 6000 (Illumina)