Hello all:

This is my first time working with lentivirus and transductions. I am trying to generate KD HCT-116 cell lines using shRNA in pTET-On vector. Can anyone answer me the following questions:

1. How does the cells look during and after transduction?

2. What should be expected cell death during transduction?

3. What are the possible reasons for higher cell death during this process?

4. what is the optimum cell confluency and cell count( for seeding)?

5. Is pTet a suitable vector for transfection and transduction?

Thanks in advance. Your answers would be of great help!!

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