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使用基于循环 RT-PCR 的策略对玉米 Mitochondrion 中初级和已处理转录的区分和映射
JoVE Journal
Genetica
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JoVE Journal Genetica
Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy
DOI:

07:26 min

July 29, 2019

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Capitoli

  • 00:04Titolo
  • 00:39Preparation of Crude Mitochondrion from Maize Developing Kernels
  • 01:45RNA 5’ Polyphosphatase Treatment and Circularization
  • 02:33Reverse Transcription and Normalization
  • 03:30PCR Amplification and Determination of Transcript Termini
  • 04:34Verification of the cRT-PCR Mapping Results by RNA Gel Blot Hybridization
  • 05:00Discrimination of Primary and Processed 5’ Ends
  • 05:26Determination of Maize mRNA Termini Using the Circular RT-PCR-based Strategy
  • 07:05Conclusion

Summary

Traduzione automatica

通过结合循环RT-PCR、定量RT-PCR、RNA 5'聚磷酸酶处理和北方斑点,提出了基于RT-PCR的循环RT-PCR策略。该协议包括一个标准化步骤,以尽量减少不稳定5'三磷酸酯的影响,它适用于对玉米线粒体中稳定积累的初级和加工转录转录进行鉴别和映射。

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