What steps would I take to obtain a population of trypsinized CHO cells to analyze using...
What steps would I take to obtain a population of trypsinized CHO cells to analyze using the haemocytometer ?
What was historically a significant challenge for making PCR successful/doable based on the use of the specific reagents required in a PCR reaction ?
Homework Answers
To answer the first question of trypsinization of CHO cells, steps are as follows:
1. Remove the medium.
2. Wash the cells twice in PBS.
3. Add Trypsin EDTA over the monolayer in a quantity enough to cover it.
4. Incubate in 37 deg Celsius for 2 minutes.
5. The cells can be checked to being dislodged from the bottom of the dish.
6. Add serum-containing medium to the cells to inhibit further trypsinization.
7. These cells can be centrifuged to obtain the cell pellet.
8. Resuspend the pellet with culture medium.
9. Take a drop of this suspension of cells over the hemocytometer and put coverslip.
10. Analyse under the microscope.
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