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PolyFast Transfection Reagent
PolyFast 轉(zhuǎn)染試劑
MCE PolyFast Transfection Reagent 采用非脂質(zhì)體陽離子聚合物為主要成分，可高效地對 DNA、RNA 進行轉(zhuǎn)染。
Description& Advantages
MCE PolyFast Transfection Reagent consists of cationic polymers and can introduce nucleic acids (DNA or RNA) into eukaryotic cells. In polymer-based transfection, exogenous DNA forms complexes with cationic polymers that enter host cells by endocytosis. PolyFast Transfection Reagent has high-efficiency, low-toxicity transfection of many cell types, including some hard-to-transfect cells.
Publications
Storage
Store at -20°C for 2 years. Avoid repetitive freeze-thaw cycles.
Protocol
1. Prepare cells
1.1 Inoculate cells in advance until the desnity reaches 70-90% for cell transfection.
Note: The viability and general health of cells prior to transfection significantly affect transfection result. Cells should be at least 90% viable prior to transfection and have had sufficient time to recover from passaging.
1.2 Remove the medium. Wash twice with PBS, and then add 900 μL serum-free medium to each well of 6-well plate.
2. Prepare PolyFast Transfection Reagent/DNA complex
2.1 According to table 1, dilute 3 μL PolyFast Transfection Reagent with 50 μL serum-free medium for each well of 6-well plate and mix gently. Incubate at room temperature for 5 minutes.
2.2 According to table 1, dilute 1 μg DNA with 50 μL serum-free medium for each well of 6-well plate and mix gently. Incubate at room temperature for 5 minutes.
2.3 Mix the diluted PolyFast Transfection Reagent and DNA gently. Incubate at room temperature for 15 minutes.
3. Add PolyFast Transfection Reagent/DNA complex to cells
Add the PolyFast Transfection Reagent/DNA complex to cells in 6-well plate and mix well. Incubate at 37°C for 24-48 hours. The medium can be replaced with fresh serum-containing medium after 6 hours if necessary.
| Table 1. PolyFast Transfection Reagent: DNA Ratio. | |||||||
| Plate Size | 96-Well | 24-Well | 12-Well | 6-Well | 60 mm | 100 mm | |
|---|---|---|---|---|---|---|---|
| Growth Area (per well) | 0.3 cm2 | 2 cm2 | 4 cm2 | 9.5 cm2 | 20 cm2 | 60 cm2 | |
| Serum-free medium | 80 μL | 450 μL | 630 μL | 900 μL | 2.7 mL | 5.4 mL | |
| Dilution of Transfection Reagent | Transfection Reagent | 0.75 μL | 1.5 μL | 2.25 μL | 3 μL | 7.5 μL | 15 μL |
| Serum-free medium | 10 μL | 25 μL | 35 μL | 50 μL | 150 μL | 300 μL | |
| Dilution of DNA or RNA | RNA | 0.25 μg | 0.5 μg | 0.75 μg | 1 μg | 2.5 μg | 5 μg |
| Serum-free medium | 10 μL | 25 μL | 35 μL | 50 μL | 150 μL | 300 μL | |
| Total Volume (per well) | 100 μL | 500 μL | 700 μL | 1 mL | 3 mL | 6 mL | |
Note: 1) Generally, the ratio of DNA (μg) to PolyFast Transfection Reagent (μL) is 1:3, and the transfection effect can be optimized in the range of 1:1 to 1:5 if necessary.
2) RNA transfection just follows the protocol as described for DNA.
4. Detection
Measure transfection efficiency using an appropriate assay for the reporter gene. For transient transfection, cells are typically assayed 24–48 hours after transfection.
PolyFast Transfection Reagent
PolyFast 轉(zhuǎn)染試劑
MCE PolyFast Transfection Reagent 采用非脂質(zhì)體陽離子聚合物為主要成分，可高效地對 DNA、RNA 進行轉(zhuǎn)染。
Description& Advantages
MCE PolyFast Transfection Reagent consists of cationic polymers and can introduce nucleic acids (DNA or RNA) into eukaryotic cells. In polymer-based transfection, exogenous DNA forms complexes with cationic polymers that enter host cells by endocytosis. PolyFast Transfection Reagent has high-efficiency, low-toxicity transfection of many cell types, including some hard-to-transfect cells.
Publications
Storage
Store at -20°C for 2 years. Avoid repetitive freeze-thaw cycles.
Protocol
1. Prepare cells
1.1 Inoculate cells in advance until the desnity reaches 70-90% for cell transfection.
Note: The viability and general health of cells prior to transfection significantly affect transfection result. Cells should be at least 90% viable prior to transfection and have had sufficient time to recover from passaging.
1.2 Remove the medium. Wash twice with PBS, and then add 900 μL serum-free medium to each well of 6-well plate.
2. Prepare PolyFast Transfection Reagent/DNA complex
2.1 According to table 1, dilute 3 μL PolyFast Transfection Reagent with 50 μL serum-free medium for each well of 6-well plate and mix gently. Incubate at room temperature for 5 minutes.
2.2 According to table 1, dilute 1 μg DNA with 50 μL serum-free medium for each well of 6-well plate and mix gently. Incubate at room temperature for 5 minutes.
2.3 Mix the diluted PolyFast Transfection Reagent and DNA gently. Incubate at room temperature for 15 minutes.
3. Add PolyFast Transfection Reagent/DNA complex to cells
Add the PolyFast Transfection Reagent/DNA complex to cells in 6-well plate and mix well. Incubate at 37°C for 24-48 hours. The medium can be replaced with fresh serum-containing medium after 6 hours if necessary.
| Table 1. PolyFast Transfection Reagent: DNA Ratio. | |||||||
| Plate Size | 96-Well | 24-Well | 12-Well | 6-Well | 60 mm | 100 mm | |
|---|---|---|---|---|---|---|---|
| Growth Area (per well) | 0.3 cm2 | 2 cm2 | 4 cm2 | 9.5 cm2 | 20 cm2 | 60 cm2 | |
| Serum-free medium | 80 μL | 450 μL | 630 μL | 900 μL | 2.7 mL | 5.4 mL | |
| Dilution of Transfection Reagent | Transfection Reagent | 0.75 μL | 1.5 μL | 2.25 μL | 3 μL | 7.5 μL | 15 μL |
| Serum-free medium | 10 μL | 25 μL | 35 μL | 50 μL | 150 μL | 300 μL | |
| Dilution of DNA or RNA | RNA | 0.25 μg | 0.5 μg | 0.75 μg | 1 μg | 2.5 μg | 5 μg |
| Serum-free medium | 10 μL | 25 μL | 35 μL | 50 μL | 150 μL | 300 μL | |
| Total Volume (per well) | 100 μL | 500 μL | 700 μL | 1 mL | 3 mL | 6 mL | |
Note: 1) Generally, the ratio of DNA (μg) to PolyFast Transfection Reagent (μL) is 1:3, and the transfection effect can be optimized in the range of 1:1 to 1:5 if necessary.
2) RNA transfection just follows the protocol as described for DNA.
4. Detection
Measure transfection efficiency using an appropriate assay for the reporter gene. For transient transfection, cells are typically assayed 24–48 hours after transfection.
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