attached is a video tutorial on Pulse chase

Thanks so much for the video,
I think that this technique will be a combination of a biochemical tool as well as cellular.
Cellular tool because:
- through this method we are going to examine an actual "cellular process (for example a secretory pathway)" and how is it occurring in a period of time in the cells.
- We are studying the activity of certain cells over a prolonged period of time.
Biochemical tool because:
- In this technique radioactive amino acids are used. A labeled compound (pulse) and then to the same compound in an unlabeled form (chase).

How to use this technique in my virus group (ssDNA viruses)?
- I think that this technique can be use in ssDNA to visualized the intranuclear distribution of parts of the virus during replication and transcription in infected cells.
- To see if viral replication occurs in multiple sites or if it is highly organized in the nucleoplasm.
- It will allowed to observed if newly synthesized viral double-stranded DNA molecules are displaced from the replication site, and how they are spread throughout the nucleoplasm.
- To see if single-stranded DNA replication is localized (in the same site) or if it accumulates in adjacent sites.
- This technique could go together with microscopy to see if replication occurs in a localized site and exactly where.
- It will help as well to visualized if sites where replication and transcriptions occurred are the same, or if they occurred in different times and how long takes the transition between this two process.
- This technique would help to determine if splicing of viral RNAs occurs co-transcriptionally and if sites of viral DNA replication are spatially distinct from the sites of RNA transcription.

Based on the information provided I would use a pulse chase analysis in my experiments to detect and analyze the path of transcripts and protein products in an in vitro or in vivo experiment using for instance wheat germ extracts or protoplasts or the TNT promega kit that are going to contain a construct with the viral genome of a pararetrovirus (dsDNA). Specifically, in my experiment I would add methionine labeled proteins or the radioactive leucine to the in vitro medium, afterwards the labeled proteins will join to the viral proteins of interest that are being synthesized. Then, the reaction will be stopped sequentially which means at different times. Additionally, in order to confirm the fusion of the tags or partners to the molecules of interest, a gel confirmation through Northern or Western blot should be performed. Parallely, a photographic emulsion would be added to the medium so that the labeled proteins that are fused with the targets can be recorded by using electron microscopy. In that way, these procedure will give as an idea of the movement or path of the molecules of interest in this virus, in my case when analysing the experiment, the transcription mechanism should be higlhy focused in the nucleus and the translation should be recorded at viroplasm level in the cytoplasm.

Thanks.