title: GSM1257963: Liver DX5+ rep2; Mus musculus; OTHER
library_construction_protocol: Spleen, liver and bone marrow cells were isolated from Rag1-/- mice and sorted for CD3-CD19-NK1.1+CD49a+DX5- or CD3-CD19-NK1.1+CD49a-DX5+ which were lysed. mRNA was extracted from cell lysates using oligo-dT beads (Invitrogen). For cDNA synthesis, we used custom oligo-dT primer with a barcode and adapter-linker sequence (CCTACACGACGCTCTTCCGATCT—XXXXXXXX-T15). After first strand synthesis samples were pooled together based on Actb qPCR values and RNA-DNA hybrids were degraded using consecutive acid-alkali treatment. Then, second sequencing linker (AGATCGGAAGAGCACACGTCTG) was ligated using T4 ligase (New EnglandBiolabs, Ipswich, MA) and after SPRI clean-up, mixture was PCR enriched 14 cycles andSPRI purified to yield final strand specific 3'end RNA-seq libraries
attributes.tag: GEO Accession
attributes.value: GSM1257963