Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy

Yafeng Zhang; Xun Liao; Peihua Luo; Kui Peng; Wenyi Ye; Tengxiang Lian; Qibin Ma; Hai Nian

doi:10.3791/60019

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Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy

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JoVE 杂志遗传学

Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy

6,034 Views

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07:26 min

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July 29, 2019

DOI:

10.3791/60019-v

DOI:

10.3791/60019-v

•

07:26 min

July 29, 2019

•

2 Views

Yafeng Zhang^1,2, Xun Liao², Peihua Luo², Kui Peng², Wenyi Ye², Tengxiang Lian^1,2, Qibin Ma^1,2, Hai Nian^1,2

¹State Key Laboratory of Conservation and Utilization of Subtropical Agro-Bioresources,South China Agricultural University, ²Guangdong Provincial Key Laboratory of Plant Molecular Breeding, College of Agriculture,South China Agricultural University

Summary

We present a circular RT-PCR-based strategy by combining circular RT-PCR, quantitative RT-PCR, RNA 5' polyphosphatase-treatment, and Northern blot. This protocol includes a normalization step to minimize the influence of unstable 5' triphosphate, and it is suitable for discriminating and mapping the primary and processed transcripts stably accumulated in maize mitochondrion.

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Mitochondrion Maize Primary Transcripts Processed Transcripts Circular RT-PCR RNA Normalization Five Prime Polyphosphatase Treatment RNA Extraction Reverse Transcription PCR

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Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy

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