First, 0.5 μg total RNA was Cy3- and Cy5-labeled with the miRCURY LNA miRNA Array labeling kit (Exiqon).
Array labeled displays an image corrupted by salt and pepper noise.
Array labeled depicts the current processing window and a pepper noise pixel.
Briefly, RNA from each sample was labeled with Cy3 using miRCURY™ LNA miRNA Array labeling kit.
Five micrograms of sample RNA were directly labeled with either Hy3 or Hy5 using the miRCURY LNA array labeling kit (Exiqon).
This was not unexpected because center 2 used the original Affymetrix array labeling protocol incorporating two labeled NTPs (biotin UTP and biotin CTP), and all other centers used a newer labeling protocol that incorporates one NTP (biotin UTP).
The same RNA used for tiling array labeling and hybridization was used as the template for qRT-PCR.
Then, the isolated miRNAs were labeled with Hy3 using the miRCURY array labeling kit (Exiqon, Vedbaek, Denmark) and hybridized to a miRCURY LNA microRNA array (version 9.2, Exiqon).
Total RNA was labelled with Hy-dye using Exiqon's miRCURY LNA array labelling kit (Vedbæk, Denmark) with the inclusion of the miRCURY LNA array synthetic spike-in miRNAs in the labelling reaction.
RNA was extracted with Trizol (Life Technologies, Carlsbad, CA, USA), subjected to quality control using a Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA), and labelled with Hy3 using the miRCURY™ LNA microRNA array labelling kit (Exiqon).
Total RNA (5 μg) was labelled by Hy3 fluorescence using a miRURYTM Array Labelling kit (Exiqon, Vedbaek, Denmark) and then concentrated using an RNeasy Mini Kit (Qiagen, Valencia, CA, USA).
