title: GSM3386132: ME4405; Homo sapiens; ncRNA-Seq
library_construction_protocol: Total RNA was extracted by ISOLATE II RNA Mini Kit (Bioline) and 20 ul aliquots of RNA were dried by vacuum concentration for 30 min at room temperature in the presence of RNAstable (Biomatrica 93221-001) After extracting the total RNA from samples, mRNA and non-coding RNAs are enriched by removing rRNA from the total RNA with kit. By using the fragmentation buffer, the mRNAs and non-coding RNAs are fragmented into short fragments(about 200~500nt), then the first-strand cDNA is synthesized by random hexamer-primer using the fragments as templates, and dTTP is substituted by dUTP during the synthesis of the second strand. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters, then the second strand is degraded using UNG(Uracil-N-Glycosylase) finally. After agarose gel electrophoresis, the suitable fragments are selected for the PCR amplification as templates. During the QC steps, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library. At last, the library could be sequenced using Illumina HiSeq 2000.
attributes.tag: GEO Accession
attributes.value: GSM3386132