(1) Q: What is the difference between DNA transfection and siRNA transfection?
A: 1. Since there are only 20 base pairs, siRNA is much smaller than DNA with thousands or even more than 10,000 base pairs.
2. The transfection reagents and methods used to transfect DNA are not suitable for transfection of siRNA.
3. siRNA transfection has stricter requirements on the cytotoxicity of transfection reagents than DNA transfection.
(2) Q: Does the transfection reagent need to be changed after transfection?
A: For the medium change, two situations can be distinguished:
1. If there is no medium change before transfection, the medium should be changed about 6 hours after transfection to ensure the nutrients needed for cell growth;
2. If there is a change before transfection The medium can be changed as usual when the medium is nutrient deficient.
(3) Q: What is the efficiency of PEI transfection reagent for transfecting siRNA and plasmid?
A: This PEI transfection reagent is specially developed for siRNA, and plasmid transfection is not recommended.
(4) Q: Can the transfection reagent be frozen?
A: Do not freeze, because the transfection reagent is a PEI cationic transfection reagent. Freezing at low temperature will destroy the activity of the PEI transfection reagent. Therefore, it is best to store it at 4 degrees to maintain the best transfection efficiency.
(5) Q: How to improve the transfection efficiency?
A:
a: Density of cells at the time of transfection, 30%-50%.
b: Opti-MEM serum-free medium should be used for siRNA and liposome dilution during transfection.
c: The medium can be changed 4-6h after transfection.
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