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A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

2.7K Views

08:53 min

September 15th, 2021

DOI :

10.3791/63014-v

September 15th, 2021


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Cellular Assay

Chapters in this video

0:04

Introduction

0:31

Cotransfection of HeLa Cells with the Reporter and U1 Small Nuclear RNA (snRNA) Plasmids

2:00

32P-Labeling of Oligonucleotides

2:52

Analysis of the Spliced Reporter Transcripts by Primer Extension

3:53

Analysis of the Spliced Reporter Transcripts by Fluorescent RT-PCR

5:08

Analysis of Variant U1 snRNA Expression by RT-qPCR

5:38

Setup and Running of Urea-PAGE Gels

6:27

Results: Reporter Based Splicing Efficiency of U1 snRNA Constructs

8:09

Conclusion

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