title: GSM1904073: E14_H33_K23_AcK_R1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted using Qiagen RNEasy Mini Kit Protocol Library construction and sequencing was performed by Bejing Genomics Institute (http://www.genomics.cn/en/index) The total RNA samples are first treated with DNase I. Then the mRNA is enriched by using the oligo(dT) magnetic beads (for  eukaryotes) or by removing rRNAs from the total RNA (for prokaryotes). 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.
attributes.tag: GEO Accession
attributes.value: GSM1904073