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  4. Cell Transfection
  5. PolyFast Transfection Reagent

PolyFast Transfection Reagent 

Cat. No.: HY-K1014
Manual SDS

MCE PolyFast Transfection Reagent consists of cationic polymers and can introduce nucleic acids (DNA or RNA) into eukaryotic cells efficiently, including some hard-to-transfect cells.

PolyFast Transfection Reagent
Size Price Stock Quantity
Free Sample (100 μL)   Apply Now  
750 μL USD 150 In-stock
1.5 mL USD 255 In-stock
7.5 mL USD 985 In-stock

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  • Description

  • Storage

  • Protocol

  • Components

  • Documentation

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.

Components
Components HY-K1014-750 μL HY-K1014-1.5 mL HY-K1014-7.5 mL
PolyFast Transfection Reagent 750 μL 1.5 mL 5×1.5 mL
Documentation
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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PolyFast Transfection Reagent
Cat. No.:
HY-K1014
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