I am trying to set up an experiment with a tightly synchronized P. falciparum culture to detect changes in expression levels of genes that are present during a particular stage of the parasite life cycle. I have attempted sorbitol synchronization and Percoll synchronization (and a combination of these two methods) and so far I've had no luck. My culture is either not synchronized enough, or I've manipulated it too much and inadvertently killed it.
Does anyone have a good synchronization protocol that produces a highly synchronized culture without killing off the culture? Any help would be immensely appreciated!
Thank you!
In fact it depends on the stage your are willing to target using a P falciparum reference strain. Sorbitol 5% for 10 minutes @ 37C then vortex 30 seconds should be fine. You can re start a new culture with fresh RBCs and culture media, then you can expect new R1 in one more day. Using percoll is for schizonts, but the parasites get damaged and do not look well. Which strains are you using?, 3d7 sould be ok, but I prefer W2. Hope I helped. If not just let me know!
Danielle Snider you need to know your Pf strain strain very well. We usually isolate Schizonts and wait for then to burst, we are currently working with 0-3 hr ring stages. You also need a a very good Giemsa stain to be able to detect such early stages.
Hi, you can use the following article for a detailed method.
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