Figuring Out ATAC-Seq
Looking at labwork and sequencing options
Previously, I hashed out a plan that involved doing ATAC-Seq with C. gigas tissues. Shelly helped me get some answers on what ATAC-Seq entails from some experts, and put those resources in this issue. The consensus is that:
- frozen tissue is difficult to work with
- the protocol will need to be optimized several times for different tissue types
- it would be easier to optimize a protocol with live tissue
I shadowed Mac today when she worked on her embryo trial! Since there’s a potential that I can do scATAC-Seq, I would need to learn how to dissociate cells. In talking to her, it seems like I’ll need to isolate nuclei no matter what I end up doing, so that’s a better method to look into first. Mac send me a nuclei isolation protocol that I’ll review and figure out how to translate to frozen C. gigas tissues.
Going forward
- Determine what kits to use for RNA and DNA extractions and order necessary materials
- Test RNA extraction protocol with tissue in histology blocks
- Start processing frozen tissues
- Extract DNA and RNA from larvae
- Identify an ATAC-Seq protocol and start testing it
- Figure out what to do with C. virginica sperm and other potential samples
Written on January 29, 2020