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ACT1-CUP1 Assays Determine the Substrate-Specific Sensitivities of Spliceosomal Mutants in Budding Yeast
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JoVE Journal Genética
ACT1-CUP1 Assays Determine the Substrate-Specific Sensitivities of Spliceosomal Mutants in Budding Yeast

ACT1-CUP1 Assays Determine the Substrate-Specific Sensitivities of Spliceosomal Mutants in Budding Yeast

DOI:
10.3791/63232-v

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07:31 min

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June 30, 2022

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1College of Arts and Sciences,Northwest University

Capítulos

  • 00:05Introduction
  • 00:47Pouring Plates for the ACT1-CUP1 Assay
  • 02:13ACT1-CUP1 Plate Preparation
  • 03:26Plating the Strains on the Copper Plates, Data Collection, and Analysis
  • 05:09Results: Impact of Mutant Splicing Factors on the Spliceosomal Function
  • 06:50Conclusion

Summary

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The ACT1-CUP1 assay, a copper growth assay, provides a quick readout of precursor messenger RNA (pre-mRNA) splicing and the impact mutant splicing factors have on spliceosomal function. This study provides a protocol and highlights the customization possible to address the splicing question of interest.

Tags

ACT1-CUP1 AssayPre-messenger RNA SplicingSplicing Factor MutationGrowth PhenotypeLeucine Dropout Growth MediaCopper SulfateOptical DensitySpectrophotometerDilutionBunsen BurnerReplicator

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