GSM7908654: Brain_Poly IC_M_repl1; Mus musculus; RNA-Seq
5 days following the Poly I:C injection on ED17.5, pregnant dams were euthanized using CO2. The uterine horn was dissected out and placenta and fetal brains were taken from each of the offspring and flash frozen with dry ice. RNA was isolated from fetal brain (n = 32; 16 males and 16 females) and placentas (n = 32; 16 males and 16 females) from 7-8 litters using RNA/DNA Miniprep Plus Zymogen Kit (Cat#D7003). To ensure purity, RNA was further treated with DNAse1 for 15 minutes. RNA quantification was done using a Qubit. Chemical purity was determined using a Nanodrop, where the absorbance ratios fell between 1.6 and 2.1(260/280) and above 1.3 (260/230). cDNA libraries were prepared according to the Lexogen manual, using 500ng of total input RNA per sample and 12 cycles of PCR amplification. 64 unique, dual index barcodes were used and samples were then pooled. QuantSeq 3'mRNA sequencing FWD (RNAseq) libraries (Lexogen) were prepared as stated in the Lexogen manual using 500ng of total input RNA per sample and 12 cycles of PCR amplification. 64 unique, dual index barcodes were used and samples were then pooled. Unique molecular identifiers were used in place of SS1. Pooled samples were exonuclease VII treated and sequenced on the HiSeq4000 to generate Single End 100 base pair reads.