Preparation of Streptavidin Coated Magnetic Beads — I
3:52
Preparation of Cell Lysate
5:36
Preparation of Streptavidin Magnetic Beads — II
6:34
Pull-down of Target mRNA-miRNA Complexes
7:35
Total RNA Extracted, cDNA Synthesis, and qPCR
8:57
Results: Use of MicroRNAs to Capture the Target mRNA
9:55
Conclusion
副本
This method can help us answer key questions in the field of mRNA biology such as, whether in silico predicted mRNA target can directly interact with a particular cellular microRNA. The main advantage of this method is that it utilizes biotinylate
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This report describes a fast and reliable method for validating mRNA targets of cellular miRNAs. The method uses synthetic biotinylated Locked Nucleic Acid (LNA)-based miRNA mimics to capture target mRNA. Subsequently, streptavidin-coated magnetic beads are employed to pulldown the target mRNA for quantification by qPCR polymerase chain reaction.
