Strain: T65H translocation mouse (Beechey et al., Mammalian Genome 8(4):236-240, 1997) without uniparental duplications (normal) Age: 13.5dpc Tissue: embryo
Biomaterial provider
Colin Beechey, Mammalian Genetics Unit, Harwell, Oxfordshire, UK
Treatment protocol
Tissues were snap frozen in liquid nitrogen, permanently stored at -80C, and put on dry ice during transport.
Growth protocol
N/A
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated from tissues in guanidine isothiocyanate by centrifugation through caesium chloride.
Label
Biotin
Label protocol
Caesium chloride isolated total RNA was quantified on an Agilent Bioanalyser and 5 to 7µg were used to prepare biotin-labeled cRNA target, as prescribed in the Affymetrix expression manual (Affymetrix, 2004). The biotinylated cRNA target was purified using cRNA Cleanup spin columns (Affymetrix), fragmented with 5x fragmentation buffer (Affymetrix), and quantified prior to hybridization on an Agilent Bioanalyser.
Hybridization protocol
0.15µg of labeled probe was hybridised to the array, washed and stained on the Affymetrix fluidics station 450. Standard protocol.
Scan protocol
The Affymetrix Scanner 3000 system was used. Standard protocol.
Description
Tissue-specific comparison of gene expression levels in T65H translocation mice, either with or without uniparental duplications of Chrs 7 & 11. Identification of highly differentially expressed transcripts. See T65H series record.
