Dataset features

Specifications


Dataset type: Other
Number of samples: 10
Release date: May 3 2016
Last update date: Apr 24 2017
Access: Public
Dataset link Early life stress perturbs the maturation of microglia in the developing hippocampus [P14]

Experimental Protocol


microglia were isolated from the hippocampus of 14-day old control and BDS male pups (n=5 from each group, all from different litters) using Percoll gradient followed by FACS analysis. RNA was harvested using the he mirVana mirRNA isolation kit (Cat # AM 1560, Life technologies) and was then concentrated and Dnase treated on column using the RNA clean and contractor-5 kit (Cat# R1015, Zymo Research). Two ng of RNA were converted to cDNA using Superscript Vilo (Cat# 11754, Life Technologies), amplified with 5 rounds of PCR according to the nCounter single cell gene expression protocol and hybridized to the mouse immune panel (Cat# 150761, NanoString Technologies), and processed according to manufacturer instructions. Counts were normalized to positive controls and 5 housekeeping genes (e.g. Alas1, Gapdh, Oaz1, Polr1b, Sdha, Tbp) that were not affected by either BDS or age and showed a coefficient of variation of less than 20%. Genes were considered “present” if at least 40% of the samples showed hybridization signal that was 2 s.d. above the mean value for the negative control probes

Repositories


GEO

GSE81036

BioProject

PRJNA320323

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Contact


Arie Kaffman