GSM4396441: T01 [3 dpi]; Physcomitrella patens; RNA-Seq
The total RNA of Mock plants, 0.5, 1, 2, and 3 dpi were extracted using TAKARA Trizol Reagent according to the protocol of manufacture. The total RNA was treated with RNase-free DNase I to eliminate DNA contamination.The enrichment of mRNA was performed with Oligo (dT)-attached magnetic beads,and fragmentation was performed using divalent cations under high temperature in NEBNext First Strand Synthesis Reaction Buffer (5×) to randomly interrupt mRNA. The first-strand cDNA chain was synthesized by random hexamers and M-MuLV Reverse Transcriptase (RNase H) with the mRNA template. Subsequently, the second-strand cDNA chain was synthesized by adding buffer, dNTPs, RNase H and DNA polymerase I. The the double stranded cDNA was purified using AMPure XP beads. Following end-pair and single nucleotide A (adenine) addition for the purified cDNA, adapters were used to distinguish the different samples by selection with AMPure XP beads for PCR amplification, following which the cDNA library was obtained.
GEO
GSM4396441