Hey Guys
I am working on HeLa cells and I need to do a clonogenic assay. Following are the steps I follow for seeding
1. Remove media and wash with 1XPBS
2. Add trypsin and incubate for 1 minute
3. Add complete media (2-3 times that of trypsin used)
4. transfer cells into 15 ml falcon tube and centrifuge for 7 minutes under 1200rpm
5. remove the suspension
6. Add 1 ml complete media and resuspend until completely homogenized
7. transfer 200 microlitres to an eppendrof and add same amount of trypan blue then count the cells and calculate the seeding volume required
Some times I take lot of time to do the 7th step. Does the cells get adhered to the 15ml falcon tube during that interval? If so how to avoid it ?
Please suggest an effective strategy
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