| Background | FectinMore™ Transfection Reagent is a low cell toxicity and non-liposome reagent optimized for DNA delivery. It provides effectively, reproducibly, and affordable benefits for scientific research. |
| Application | Cell Culture |
| Stability & Storage | -20℃ |
| Shipping | Blue ice |
(1) Cell preparation: Cells should be seeded before 16 to 20 hours prior to transfection with around 70% confluence. The medium should be refreshed 30 minutes before transfection. Usually, culture medium with serum does not affect transfection.
(2) DNA preparation: DNA plasmid for transfection should be with high purity (A260/A280=1.8-1.9) to ensure efficient transfection mixture preparation.
(3) Mixture preparation: Guideline of DNA plasmid and FectinMore™ DNA Transfection Reagent amount and ratio refer to table 1. In brief, plasmid and transfection reagent are diluted in serum-free culture medium for 5 minutes then mixed together and tip gently for another 25 minutes.
(4) Transfection: Add mixtures into cell culture dish/plate. The mixture could be removed after 6 to 48 hours and refilled with culture medium.
Recommended formula of transfection mixture：
| Culture Dish/Plate | Medium Volume | Plasmid / Serum-free medium |
FectinMore™ DNA Transfection Reagent / Serum-free medium |
|
96-well |
100 μ L |
250 ng/10 μ L |
0.75 μ L/10 μL |
|
24-well |
500 μ L |
500 ng/25 μ L |
1.5 μ L/25 μ L |
|
12-well |
700 μ L |
750 ng/35 μ L |
2.25 μ L/35 μ L |
|
6-well |
1 m L |
1 μg/50 μ L |
3 μ L/50 μL |
|
60mm dish |
3 m L |
2. 5 μg/150 μ L |
7. 5 μ L/150 μL |
|
100mm dish |
6 m L |
5 μg/300 μ L |
15 μ L/300 μL |
Important notes
FectinMore™ DNA Transfection Reagent is recommended for use with Gibco™ Opti-MEM™ I Reduced-Serum Medium or Gibco™ OptiPRO™ SFM for alternative optimizing transfection efficiency.
|
· Transfected with FectinMore™( FectinMore™：DNA=3：1) ，detect the expression of target protein after 2 days. |
|
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Proteins expression checked by western blot in specific primary antibody followed by secondary antibody (anti-Mouse IgG(H+L)- HRP). |
 |
Proteins expression checked by FACS in specific primary antibody followed by secondary antibody (anti - Mouse IgG(H +L)- FAM). |
 |
Proteins expression checked by immunofluorescence (IFA) in specific primary antibody followed by secondary antibody (anti - Mouse IgG -iFluor 594). |
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