GSM8654777: Wildtype_14DAP_Biologicalrep2; Thlaspi arvense; RNA-Seq
RNA was extracted using the CTAB method The mRNA was enriched and purified using oligo dT beads to select for poly(A)-tailed mRNA. RNA fragmentation was followed by first-strand cDNA synthesis using random N6 primers, and second-strand cDNA synthesis was subsequently performed. The synthesized cDNA was subjected to end-repair, followed by 3' adenylation, and adaptors were ligated to the 3' adenylated cDNA fragments. Multiple rounds of PCR amplification were then carried out to enrich the cDNA template using appropriate primers. The PCR products were denatured by heat, and cyclization was achieved through splint oligos and DNA ligase. DNA nanoballs were synthesized from the cyclized products, and sequencing was performed on the DNBSEQ platform (DNBSEQ Technology).