Select the Sequence to be Cloned in an Expression Vector
3:35
Design Primers for Cloning
4:59
Cloning of Constructs
7:05
Transfection of the Reporter Genes into Eukaryotic Cells
10:21
Results: Analysis of Circular RNAs
11:09
Conclusion
Transcription
This protocol is significant because it allows the user to generate a genomic expression system that can undergo alternative splicing and in this case form circular RNAs that can be tested for their function and disease association. The main advan
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We clone and analyze reporter genes generating circular RNAs. These reporter genes are larger than constructs to analyze linear splicing and contain Alu elements. To investigate the circular RNAs, the constructs are transfected into cells and resulting RNA is analyzed using RT-PCR after removal of linear RNA.