We tried several times to isolate RNA from HK-2 cells with the qiagen RNeasy kit (with RLT buffer). We can do the isolation of RNA (with Qiagen kit) from other lines of human immortalized proximal tubular epithelial cells without any problem, but from the HK-2 cells it's very hard to get RNA. We already tried to homogenize the sample better, using needle and syringe. We also already added the RLT buffer immediately to the culture dish (instead of to the pellet) and scraped the cells. All without improvement...
What else could we try?
Try trizol method, for reference see
Chomczynski P1, Sacchi N. The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: twenty-something years on. Nat Protoc. 2006;1(2):581-5.
Use the trizol method as Dr. Murthy indicates. There are several commercial denominations but all are the same product essentially. I used it some time ago with HK-2 cells and didn't have any problems with it.
- Badges
- Science topic
- Similar topics
- Methodology
- Laboratory
- Cellular Biology Techniques