Simple and nonradioactive detection of microRNAs using digoxigenin (DIG)-labeled probes with high sensitivity

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FIGURE 1.
FIGURE 1.

Schematic illustration of the protocol. (1) Annealing of the labeled probe, miRNA, and bridge oligonucleotide by slowly decreasing temperature after denaturation. (2) Filling in the nick with T4 DNA ligase. (3) Denaturing the double-stranded structure. (4) Fifteen percent urea–polyacrylamide gel electrophoresis. (5) Transferring the products to a membrane and cross-linking RNA by EDC. (6) Detecting the DIG-labeled miRNAs and free probes on the membrane using chemiluminescent substrates.

This Article

  1. Published in Advance February 26, 2014, doi: 10.1261/rna.042150.113 RNA 20: 580-584

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