GSM6597760: RNA-Seq data of lasermicrodissected PVN of hamster EH03_20 sampled at ZT07 on a TFD; Phodopus sungorus; RNA-Seq
20 µm kryosections were prepared in a RNAse-poor environment on Poly-L-Lysine coated Carl Zeiss™ MembraneSlides 1.0 PEN (Fisher Scientific GmbH, Schwerte, Germany) using a cryostat “Microm HM 560” (Thermo Scientific, Walldorf, Germany). The sections were kresylviolett-stained to enable lasermicrodissection of the hypothalamic core areas SCN and PVN in Carl Zeiss™ adhesive-cap tube per sample (Fisher Scientific GmbH, Schwerte, Germany) on a PALM MicroBeam (Carl Zeiss Microscopy Deutschland GmbH, Oberkochen, Germany). Total RNA was purified using the RNeasy Plus Micro Kit with gDNA eliminator spin column (Qiagen GmbH, Hilden, Germany). Total RNA was checked for quality (Bioanalyzer, Agilent Technologies, Waldbronn, Germany) and quantity (RNA 6000 Pico Kit for very small RNA amounts of 50–5000pg/µL, Agilent Technologies, Waldbronn, Germany). The mRNA were isolated, the library prepared (NEBNext® Single Cell / Low Input RNA library Prep Kit dual index), and the RNA-Seq performed on an Illumina NextSeq 500 (1 lane, 2x25 mio reads, 2 x 150 nt, 7.5 gb).