GSM1855252: ZT6-B tech rep2; Mus musculus; RNA-Seq
Brains were removed at one of six zeitgeiber times (ZT) and immediately frozen on dry ice in OCT mounting media (VWR, Lutterworth, UK). Eighteen frozen coronal sections at 15mM were cut spanning the entire rostral to caudal region of the SCN and mounted onto polyethylene napthalate (PEN) slides (Carl Zeiss Ltd. Cambridge, UK). Laser-capture microdissection was carried out from dehydrated, Nissl-stained sections using the PALM system (Carl Zeiss Ltd. Cambridge, UK) as previously described (Dulneva et al., 2015). Tissue from five animals was pooled to generate one biological replicate sample and total RNA was purified using the RNeasy micro kit (Qiagen, Manchester, UK). RNA quality was determined using an RNA Picochip (Agilent, Stockport, UK), with RIN values over 8 for all samples and average yield of approximately 10ng per replicate.  Approximately 1ng of RNA was amplified using the SMARTer protocol (Takata Clontech Bio Europe SAS, France) and the resulting libraries were 100bp paired-end sequenced on the HiSeq (Illumina, Fulbourne, UK; Wellcome Trust Centre for Human Genetics Sequencing Core).
GEO Accession
GSM1855252