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Chapters
- 00:04Title
- 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
Vi præsenterer en cirkulær RT-PCR-baseret strategi ved at kombinere cirkulær RT-PCR, kvantitativ RT-PCR, RNA 5 ' polyfosfatase-behandling og Northern blot. Denne protokol indeholder en normalisering skridt til at minimere indflydelsen af ustabile 5 ' trifosfat, og det er egnet til at diskriminere og kortlægge de primære og forarbejdede udskrifter stabilt akkumuleret i majs mitochondrion.
