Strain: C57Bl/6, Gender: female, Age: 12-14 weeks, Pooled samples: 4
Extracted molecule
polyA RNA
Extraction protocol
25,000- 50,000 cells were FACS sorted, RNA was isolated with RNAqueous Kit (Ambion) per manufacture's protocol, DNAse treated with DNAse1, phenol:chloroform:isoamyl alcohol extracted, and amplified through two rounds of in vitro transcription using MessageAmp (Ambion) per manufacture's protocol
Label
Biotin
Label protocol
Second round of MessageAmp(Ambion) amplification was biotinylated with bio-CTP and bio-UTP per manufature's protocol.
Hybridization protocol
Standard protocol from manufacturer
Scan protocol
Hardware: GeneChip¨ Scanner 3000
Description
not applicable
Data processing
Normalization was done with the GCRMA package from Bioconductor