title: GSM1412445: human smRNA-seq in HEK293T cells rep1; Homo sapiens; miRNA-Seq
library_construction_protocol: For all the in vitro dsRNA-seq and ssRNA-seq libraries, RNAs were treated with ssRNase (RNaseOne) and dsRNase (RNase V1), respectively; For mRNA-seq libraries poly A+ RNAs were isolated from total RNA using Dynabeads mRNA direct kit (Ambion, Austin, TX); For smRNA-seq libraries RNAs were prepared as previously described in F. Li et al., The Plant cell 24, 4346. Briefly, total RNA was run on a 15% TBE-Urea Gel and gel slices containing 15-35 nt RNAs were isolated and eluted from acrylimalyde by mixing with 0.3 M NaCl for 4 hours at RT. The dsRNA-seq, ssRNA-seq, mRNA-seq and smRNA-seq libraries were constructed as previously described in F. Li et al., The Plant cell 24, 4346. The 3’ and 5’ adapters, Reverse Transcriptase Primer, 5’ PCR primer, and 3’ PCR Primers matching the TruSeq small RNA sequencing kit (Illumina, San Diego, CA) were synthesized and HPLC purified by IDT (Coralville, IA).
attributes.tag: GEO Accession
attributes.value: GSM1412445