ChIP-Atlas: SRX685923

Antigen

Cell type Class: Neural
Cell type: Cerebellum
MeSH Description: The part of brain that lies behind the BRAIN STEM in the posterior base of skull (CRANIAL FOSSA, POSTERIOR). It is also known as the "little brain" with convolutions similar to those of CEREBRAL CORTEX, inner white matter, and deep cerebellar nuclei. Its function is to coordinate voluntary movements, maintain balance, and learn motor skills.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Crosslinked material was sonicated with a Bioruptor (Diagenode) with 30 sec. on/off cycles to an average size range of 150-350bp as visualized by agarose gel electrophoresis. Sonicated supernatants were diluted 10-fold in dilution buffer (0.01% SDS, 1.1% Triton X-100, 1.2mM EDTA, 16.7mM Tris pH 8.1, 167mM NaCl) before immunoprecipitation. 6uL antibody (anti-Zic 1/2 C-terminus, or total rabbit IgG, Millipore, Cat. #12-370) was first incubated with 100uL Dynabeads Protein G (Invitrogen) beads for 4 h. at 4°C, then the antibody conjugate was added to 6 mL of cell lysis for overnight IP. Standard TruSeq adapters were ligated for library preparation using the MicroPlex Library Preparation Kit (Diagenode). C57BL/6Ncrl male and female mice at P7 or P60 were deeply anesthetized with isofluorane and decapitated for brain harvesting. The cerebellar cortex was dissected, dounced in 1% formaldehyde PBS buffer, incubated at room temperature for 15 minutes, washed 2x with cold PBS, and then lysed in 600 uL lysis buffer (1% SDS, 10mM EDTA, 50mM Tris pH 8.1). We pooled cerebellum or cortex from three P7 mice or two P60 mice for each biological replicate.