A subscription to JoVE is required to view this content. Sign in or start your free trial.
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 presenterer en sirkulær RT-PCR-basert strategi ved å kombinere sirkulær RT-PCR, kvantitativ RT-PCR, RNA 5 ' polyphosphatase og Northern blot. Denne protokollen inkluderer en normalisering skritt for å minimere påvirkning av ustabile 5 ' trifosfat, og det er egnet for diskriminerende og kartlegging den primære og bearbeidede transkripsjoner stabilt akkumulert i mais mitokondrie.
