title: GSM2585282: OESC MdsEvi1 WT n2; Mus musculus; RNA-Seq
library_construction_protocol: Pregnant female mice of ICR or ICR/BL6 mixed background carrying the d2EGFP transgene under the control of the Hes1 or Hes5 promoters (Ohtsuka et al., 2006, PMID 16214363.) were sacrificed at embryonic day 12.5 (E12.5) and olfactory epithelia (OE) were isolated. Tissue was dounced 10 times and incubated in Trypsin EDTA solution at 37°C for 3 minutes to generate cell suspensions. Trypsin treatment was quenched by addition of DMEM/10% FBS and GFP-positive cells were then isolated by FACS. Sorting was performed directly into RNA isolation buffer and total RNA was extracted using the RNeasy Micro Kit (Qiagen). Library construction and sequencing were carried out as described here: 1) library preparation:   Salimullah M, Sakai M, Plessy C, Carninci P. NanoCAGE: a high-resolution technique to discover and interrogate cell transcriptomes. Cold Spring Harb Protoc. 2011 Jan 1;2011(1):pdb.prot5559. PubMed: 21205859 )  2) template switching oligo containing TATA spacer:  Tang DT, Plessy C, Salimullah M, Suzuki AM, Calligaris R, Gustincich S, Carninci P. Suppression of artifacts and barcode bias in high-throughput transcriptome analyses utilizing template switching. Nucleic Acids Res.   2013 Feb 1;41(3):e44. PubMed: 23180801  3) methodology:  Plessy C, Bertin N, Takahashi H, Simone R, Salimullah M, Lassmann T, Vitezic M, Severin J, Olivarius S, Lazarevic D, Hornig N, Orlando V, Bell I, Gao H, Dumais J, Kapranov P, Wang H, Davis CA, Gingeras TR, Kawai J, Daub CO, Hayashizaki Y, Gustincich S, Carninci P. Linking promoters to functional transcripts in small samples with nanoCAGE and CAGEscan. Nat Methods. 2010 Jul;7(7):528-34 PubMed: 20543846
attributes.tag: GEO Accession
attributes.value: GSM2585282