GSM2175156
GSM2175156: Total RNA from wild type; Metarhizium robertsii ARSEF 23; ncRNA-Seq
Metarhizium robertsii wild type and Δdcl2 were ground into powder in liquid nitrogen,powder was used to isolate total RNAs  using Trizol (Invitrogen, USA). The total RNA samples are first treated with DNase I to degrade any possible DNA contamination. Then the mRNA is enriched by using the oligo(dT) magnetic beads. Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments. Then the first strand of cDNA is synthesized by using random hexamer-primer. Buffer, dNTPs, RNase H and DNA polymerase I are added to synthesize the second strand. The double strand cDNA is purified with magnetic beads. End reparation and 3’-end single nucleotide A (adenine) addition is then performed. Finally, sequencing adaptors are ligated to the fragments. The fragments are enriched by PCR amplification. During the QC step, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used to qualify and quantify of the sample library. The library products are ready for sequencing via Illumina HiSeqTM 2000.
GEO
GSM2175156
GEO Accession
GSM2175156