title: Illumina NovaSeq 6000 paired end sequencing; Analysis of mRNA abundance in the wild-type or Phd2 knockout carotid body and adrenal medulla of mice
design: Analysis of mRNA abundance in the wild-type or Phd2 knockout carotid body and adrenal medulla of mice
library_construction_protocol: Tyrosine hydroxylase-restricted Phd2 knockout mice were generated by inter-crossing Phd2f/f and THCre strains as described by Lindeberg et al., 2004 and Mazzone et al., 2009. Carotid bodies or adrenal medullas were sub-dissected and pooled from 5 Phd2 knockout or 5 control (THCre negative or Phd2 wildtype) mice. Tissues were stored in ice-cold RNAProtect (Qiagen) until processing. Tissues stored in RNAProtect were washed with PBS, resuspended in RLT Buffer (Qiagen) supplemented with dithiothreitiol (Merck), and homogenized with Pro200 homogenizer (ProScientific). RNA was extracted from the lysate using RNeasy Plus Micro Kit (Qiagen) according to the manufacturer's instructions and as described previously by Gao et al., 2017. RNA sequencing library was prepared using the NEBNext Single Cell/Low Input RNA Library Prep kit (NEB).