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Do you have a suggested protocol for cell transfection?
Posted February 19, 2020
Answer
Your optimal protocol will depend on multiple factors including your chosen cell line, but we do have a suggested protocol for our transfection reagent.
Sample Protocol below or available here:
1. Prepare cells for transfection
- Culture cells to ~90% confluency and use fresh, clean growth medium
2. Prepare Transfectamine™ 5000-DNA mixture
- First mix 2.5 ug of DNA with 200 uL of serum-free medium.
- Add 7.5 uL of thawed Transfectamine™ 5000 to DNA/medium mixture.
- Mix well and incubate at room temperature for 20 minutes.
3. Add Transfectamine™ 5000-DNA mixture to cell culture
4. Culture overnight
5. Analyze transfection efficiency with appropriate method
- Maximum expression levels can be discerned 72-96 hours post-transfection
- If using Flow Cytometry, Laser-Scanning Molecular Imaging, Microscopy, or Western Blot Analysis, a fluorescent label is necessary or strongly recommended
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