Total RNA was extracted from kidneys using miRNeasy mini kits (Qiagen)
Label
Cy5
Label protocol
The small RNA fraction (<300 nucleotides) was hybridized on a μParaflo Microfluidic chip containing detection probes for all mouse microRNAs (miRNAs) in the miRBase version-17 (miRBase, http://microrna.sanger.ac. uk/sequences). The hybridized microarray chips were labeled with fluorescent dyes and laser scanned to obtain fluorescent images. The signal values for each sample were derived by background subtraction and normalization. Microarray chip hybridization, fluorescent labeling, laser scanning, and background subtraction and normalization were performed by LC Sciences proprietary method.
Hybridization protocol
The small RNA fraction (<300 nucleotides) was hybridized on a μParaflo Microfluidic chip containing detection probes for all mouse microRNAs (miRNAs) in the miRBase version-17 (miRBase, http://microrna.sanger.ac. uk/sequences). The hybridized microarray chips were labeled with fluorescent dyes and laser scanned to obtain fluorescent images. The signal values for each sample were derived by background subtraction and normalization. Microarray chip hybridization, fluorescent labeling, laser scanning, and background subtraction and normalization were performed by LC Sciences proprietary method.
Scan protocol
The small RNA fraction (<300 nucleotides) was hybridized on a μParaflo Microfluidic chip containing detection probes for all mouse microRNAs (miRNAs) in the miRBase version-17 (miRBase, http://microrna.sanger.ac. uk/sequences). The hybridized microarray chips were labeled with fluorescent dyes and laser scanned to obtain fluorescent images. The signal values for each sample were derived by background subtraction and normalization. Microarray chip hybridization, fluorescent labeling, laser scanning, and background subtraction and normalization were performed by LC Sciences proprietary method.
Data processing
The signal values from each of the five age groups (P2, P7, P14, P35) were averaged, and the P values using One-way ANOVA were calculated. The differentially detected signals are defined by P < 0.05.
