title: Illumina HiSeq 4000 paired end sequencing; RNA-seq of of bone marrow derived mononuclear phagocytes
design: RNA-seq of of bone marrow derived mononuclear phagocytes
library_name: rabbit3_LPS4_p
library_construction_protocol: Cells were washed with PBS and then detached using a non-enzymatic solution.  Cells were stimulated with either 100ng/mL lipopolysaccharide (LPS) (Invivogen, Cat. Code: tlrl-smlps) or with 1ug/mL of  High-Molecular Weight poly I:C (Invivogen, Cat. Code: tlrl-pic), transfected with 2uL/mL Lipofectamin 2,000, for 4 hours before collection. Total RNA was extracted using the RNeasy Plus Mini kit(Qiagen, Cat Number  74136), using QIAcube (Qiagen).  Libraries were produced using the Kapa Stranded mRNA-seq Kit (Kapa Biosystems, Kit code: KK8421). cDNA was amplified in 13 PCR cycles, and purified by Ampure XP beads (Beckman Coulter, Cat Number A63882) (1.8x volume) using Zephyr (Perkin Elmer). Prior to sequencing, libraries were quantified using MiSeq (25bp PE). This data was used for pooling libraries in equimolar amounts by Coulter NX (Span-8, Beckman).
attributes.tag: Experimental Factor: organism
attributes.value: Oryctolagus cuniculus
attributes.tag: Experimental Factor: stimulus
attributes.value: lipopolysaccharide; 100 nanogram per milliliter
attributes.tag: Experimental Factor: time
attributes.value: 4