I want to extract RNA from that cells. Does trypsin like usual culture works? Im using the classic inserts and covering them with geltrex dil 1/4 in RPMI
I have never used these plates, but could you put your phenol based rna extraction solution into the well (tryzol), it would lyse all the cells where they were.
Best wishes
If you detach your cells from the plates using trypsin, you can use that. Put trypsin in the bottom of the well and let the insert soak in the trypsin (unless is too much volume to use), then wait some minutes or even incubate it at 37ºC for a while and flush using a pipette collecting the media from the bottom and flushing the surface. I have done it that way and it worked.
I wouldn't bother with trypisinization. Your cells are attached and likely polarized at this point.
I've worked with MDCKs and retinal pigment epithelium cells in Transwell plates (polycarbonate, polyester, and even PTFE). Even 20+ minutes of trypsin with a pre-treatment of HBSS to remove tight junctions doesn't make them come off very well. The yield has always been pretty poor from a protein standpoint and honestly, it seems like I'm just doing more harm to them beforehand.
Just take a cell scraper to it with some lysis buffer. Spray the lysis buffer on and just scrape away. I don't know what size of Transwell you use, but try to go with the broadest scraper you can fit since Transwells are only a few microns thick. and you don't want to punch a hole into them and lose your sample.
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts