Chimeric RNAs can be formed by trans-splicing from different transcripts or cis-splicing of adjacent genes (cis-SAGe). Cis-SAGe results from read-through transcription of two neighbor genes. To investigate the mechanisms underlying intergenic splicing of adjacent genes, it is important to develop an assay to detect transcriptional read-through. Here, we describe a general RT-PCR based method to confirm the process for cis-SAGe candidates. In this method, we use PCR to amplify cDNA that is reverse transcribed from the read-through precursor mRNA. The result provides a foundation for further downstream mechanistic studies.
CITATION STYLE
Shi, X., Qin, F., & Li, H. (2020). Confirmation of Transcriptional Read-Through Events by RT-PCR. In Methods in Molecular Biology (Vol. 2079, pp. 177–186). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9904-0_14
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