GSM4561734: 14_S137: dHC-Naive_rep2; Rattus norvegicus; RNA-Seq
Rats were euthanized by decapitation. Their brains were quickly dissected and snap-frozen in pre-chilled isopentane on dry ice. PL (bregma +3.7mm to +2.2mm) and dHC (bregma -2.3mm to -4.52mm) tissue was isolated by punches in a cryostat using a Neuro Punch (19 gauge, Fine Science Tools, Foster City, CA) as described previously (Milekic et al., 2007) and immediately submerged into Qiazol (Qiagen, Venlo, Netherlands). Total RNA was isolated using the RNAeasy Plus Universal Kit following the manufactureâ€™s protocol (Qiagen, Venlo, Netherlands). 500 ng of RNA was reverse-transcribed using QuantiTect Reverse Transcription Kit (Qiagen, Venlo, Netherlands) in order to perform real-time polymerase chain reaction (RT-PCR) on all individual samples to validate immediate early gene c-Fos induction in trained vs. naÃ¯ve animals. RNA samples for naÃ¯ve and both groups of trained animals were pooled into 3 biological replicates (n=5/group per condition) and submitted to the New York University School of Medicine Genome Technology Center for further quality control, quantification and normalization. RNA integrity was tested on an Agilent 2100 Bioanalyzer before library preperation. RNA-Seq library preps were made using the Illumina TruSeqÂ® Stranded mRNA LT kit (Cat #RS- RS-122-2101 or RS-122-2102), on a Beckman Biomek FX instrument, using 200 ng of total RNA as input, amplified by 12 cycles of PCR, and run on an Illumina 4000, as paired end 150 reads.
GEO Accession
GSM4561734