RNA-seq profiling of IgE and IgG1 repertoire from the bone marrow of allergic mice >7 weeks post challenege.
Balb/c mice were sensitized to egg white and egg yolk intratracheally for 2 weeks and challenged with the same allergen for 7 weeks or more. After the 7th week, the femur and tibia were collected from allergic mice. The bone marrow was extracted by cutting both ends of the bones and flushing them with 20ml of PBS to obtain a single-cell suspension. The cells were then centrifuged, the supernatant was discarded, and the pellet was resuspended in RNAlater and stored at –20?°C. RNA was extracted using RNeasy Mini plus kit (Qiagen). RNA was used for the construction of the sequencing library. Libraries for RNA seq was prepared using the iRepertoire MBHI kit following the manufacturer's instructions.