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2.3K Views
•
07:31 min
June 30th, 2022
DOI :
10.3791/63232-v
Chapters
0:05
Introduction
0:47
Pouring Plates for the ACT1-CUP1 Assay
2:13
ACT1-CUP1 Plate Preparation
3:26
Plating the Strains on the Copper Plates, Data Collection, and Analysis
5:09
Results: Impact of Mutant Splicing Factors on the Spliceosomal Function
6:50
Conclusion
Transcript
ACT1-CUP1测定可以确定突变是否影响信使RNA剪接前,并指示哪个剪接步骤受影响最大。ACT1-CUP1测定将剪接因子突变与其对复杂剪接过程的影响联系起来。它利用生长表型的简单读数,具有广泛的灵敏度范围。
在进行ACT1-CUP1测定时,有效利用时间很重要。从 5 到 10 种菌株的测定开始,以尽量减少压力引起的生长畸变。演示这个过程的将是我和我实验室的本科研究员Isabelle Marasigan。
首先,向每个瓶子中加入 790 毫克琼脂和搅拌棒。在一个大烧
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Summary
ACT1-CUP1测定是一种铜生长测定,可快速读取前体信使RNA(pre-mRNA)剪接以及突变剪接因子对剪接体功能的影响。本研究提供了一个协议,并强调了解决感兴趣的拼接问题的定制可能。
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