IN order to clone a mature miRNA: What is the most efficient protocol? Thanks
Dear Devrim,
Please read the following text:
https://www.thermofisher.com/us/en/home/references/ambion-tech-support/microrna-studies/tech-notes/microrna-cloning-overview.html
MicroRNA Cloning Overview
MicroRNAs (miRNAs) are an important class of small RNAs that regulate protein translation. To date, 230 mammalian miRNAs have been identified and cataloged in the Sanger miRNA Registry. Some scientists believe that the number of miRNAs could exceed 1000 [1]. While cloning and sequencing were used to identify most of the known mammalian miRNAs, increasing effort is being put into bioinformatic prediction of new miRNA sequences. A combination of computational biology tools and molecular cloning methods will likely be used for characterizing additional miRNAs.
Isolating the mature miRNA fraction is the first critical step in cloning these small regulatory molecules. Ambion scientists have developed two sequential systems that are optimized for isolating and enriching RNA preparations that contain 17–24 nucleotide, mature miRNAs. Here we describe the process by which miRNAs can be cloned in an effort to identify new members of this small RNA family.
1. Isolate total RNA using a method that will capture small RNAs
mirVana miRNA Isolation Kit includes small RNAs in the total RNA preparation and can subsequently be used to enrich for RNA
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