Product Informations

• Product No. KX0110047
• Product Name BioShuttle PEI-293 Adherent Cell Transfection Reagent
• Category 525
• Instruction Manual /files/uploads/instructions/2024/202403/KX0110047_BioShuttlePEI-293贴壁细胞转染试剂_说明书240314.pdf

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Product Details

Usage: It is used for transient transfection of adherent cells and construction of stable cell lines.
Transportation and storage: QuickPEI transfection reagent is recommended to be transported at room temperature. Sterile use; Store at -20 ° C. Valid for 24 months. It can be stored for a long time at -80 ° C. If frozen, store at 2 to 8 ° C after thawing (can be stored for 6 months), avoiding repeated freezing and thawing, which may affect the transfection effect.
Filtration conditions: This product is sterilized by filtration through a 0.22µm filter membrane.
 
Product Description
QuickPEI transfection reagent is a compound transfection reagent free of animal-derived components and is a powerful transient transfection reagent. Provide efficient and repeatable gene delivery solutions for adherent cells. In the HEK293 and CHO expression systems, QuickPEI provides uniform high gene expression over a wide range (from 96-well plates to 100L bioreactors). It is particularly suitable for automatic pre-preparation or manual HTS (high-throughput screening) transformation protocols, and can be used for antibody expression, various virus packaging such as AAV, lentivirus, adenovirus, and reagents for recombinant proteins.
 
 
Adherent cell transfection usage method (taking 6-well plates as an example) :
1. Cell inoculation: To enhance transfection efficiency, it is recommended to inoculate cells (such as 293 cells) into a 6-well cell plate 24 hours before transfection, with 5 to 10×105 cells per well and 3ml of complete culture medium. During transfection, the cell density is preferably between 70% and 80%.
2. Prepare the DNA-PEI complex: Prepare the DNA-PEI nucleic acid-transfection reagent complex according to the following system:
1）For each well of cells, dilute 2 μg of the target DNA with 100 μL of serum-free diluent (such as DMEM, etc.) and mix thoroughly to form a DNA diluent.
【 Note 】 It is recommended to use Opti-MEM, DMEM or ddH2O for serum-free diluents, and Opti-MEM is preferred.
2) Immediately add 6 μL of QuickPEI transfection reagent to 100 μL of DNA diluent and gently mix. The ratio of DNA to PEI (mass-to-volume ratio, µg:µl) can be adjusted as needed
3）Incubate at room temperature for 10 to 15 minutes to form a DNA-cationic nucleic acid transfection reagent complex.
3. Transfection of cells
1)During the formation of the complex, remove the cell growth medium and add 2 mL of freshly preheated complete medium to each well.
2) Add all the DNA-PEI complexes to the cells, shake the culture plate and gently mix.
3) Incubate in a 37 ℃ and 5% CO2 incubator. The expression of the transfected gene can be detected as soon as 24 hours after transfection. Please determine the appropriate time for the test by yourself.
4. Stable Rotation Screening (Optional)
After transfection for 24 hours, the cells were subcultured into fresh growth medium (diluted more than 10 times), and incubated overnight at 37 ° C in a 5% CO2 incubator. The next day, add the screening drug that matches the transfection resistance gene. Drug-resistant clones can be screened out in about 1 to 2 weeks. During this period, the growth medium containing the screened drugs needs to be frequently replaced.
 
 
 
Appendix 1: QuickPEI Dosage List (Taking DNA:PEI Ratio 1:3 as an example, for reference only, optimization is recommended)

Culture vessel	Surface area (cm ²)	Medium volume (ml)	DNA dosage (μg)/DMEM (μl)	Transfection reagent dosage (μl)/DMEM (μl) dosage (μl)
96-well plate	0.4	0.2	0.1/25	0.3/25
24-well plate	2	1	0.5/50	1.5/50
12-hole plate	4	2	1/50	3/50
6-hole plate	9	3	2/100	6/100
35mm Petri dish	9	3	2/100	6/100
60mm Petri dish	21	6	4/200	12/200
100mm Petri dish	55	15	12/400	36/400
25cm ² culture flask	25	7	6/200	7.5/200
75cm ² culture flask	75	20	18/400	54/400
175cm ² culture flask	175	50	40/1000	120/1000

 

 

Partial transfection effect diagram

Transfection effect of adherent 293T cells (DNA:PEI=1 µg:3µl)


Transfection test results of 293T cells with different DNA:PEI ratios: EGFP positive plasmid, experimental 24-well plate,DNA:QuickPEI, 1:3,1:4,1:5 (DNA mass - PEI volume ratio µg:µl)


QuickPEI transfection packaged AAV virus infects 293T cells