Wild type, Sprague-Dawley rat, E13 ureteric bud, culture for 4 days
Extracted molecule
total RNA
Extraction protocol
Three ureteric buds were isolated from E13 Sprague-Dawley rats. Isolated UBs were suspended in a mixture of 50% growth factor reduced-Matrigel and 50% type 1 collagen matrix in a inner well of Transwell filter (Corning). Medium containing pleiotrophin (PTN) purified from BSN-CM (approximately 3 microgram/ml), FGF1 (250 ng/ml), and GDNF (125 ng/ml) was applied to an outer well and the culture was placed in 5% CO2 incubator at 37 C. At 4 days of culture, isolated UBs were removed from the gel. Total RNA was extracted using PicoPure RNA isolation kit (Arcturus). Double IVT was performed using MessageAmp kit (Ambion).
Label
biotin
Hybridization protocol
standard Affymetrix procedures
Scan protocol
standard Affymetrix procedures
Description
Isolated ureteric buds in three-dimensional culture exhibited non-branching growth
Data processing
Affymetrix GCOS (GeneChip Operating Software) version 1.2
