Wild type embryonic ureteric bud at E11.5 isolated by fine manual microdissection, Cd-1 strain
Extracted molecule
total RNA
Extraction protocol
Ureteric bud was isolated by treating dissected E11.5 kidney rudiments, up to T-shaped stage, with 0.5 mg/ml collagenase B (Roche, Indianapolis, IN), in D-MEM (Invitrogen, Carlsbad, CA) for thirty minutes at 37° C and carefully dissecting the mesenchyme from the UB, followed by storage at -80° C. RNA from 3 to 9 dissected ureteric buds was prepared using the RNeasy Micro Kit (Qiagen, Valencia, CA), with 30 ng poly-inosine carrier (Epicentre, Madison, WI) added to the RLT buffer. Target RNA was prepared using the TargetAmp™ 2-Round aRNA Amplification Kit 1.0 (Epicentre, Madison, WI), and hybridized to Affymetrix MOE430_v2 microarrays.
Label
biotin
Label protocol
Target RNA was prepared using the TargetAmp™ 2-Round aRNA Amplification Kit 1.0 (Epicentre, Madison, WI), and hybridized to Affymetrix MOE430_v2 microarrays.
Hybridization protocol
standard Affymetrix procedures
Scan protocol
standard Affymetrix procedures
Description
Wild type embryonic E11.5 Ureteric bud isolated fine manual microdissection replicate 3