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SureFECT™ Transfection Reagent

The SureFECT Transfection Reagent has been discontinued effective January 1, 2011. Please see the Attractene Transfection Reagent

Catalog No. Content Price
SA-01 Enough reagent for 12 SureSilencing siRNA Arrays
Or 1200 plasmid or construct transfections in 96-well plates
Or 0.5 ml

Description
The SureFECT Transfection Reagent is the high-efficiency, low-toxicity solution for the reverse transfection of a wide variety of cultured mammalian cells. Reverse transfection saves an entire day over traditional transfection by plating cells directly into wells and medium already containing reagent-DNA complexes. Reverse transfection produces equivalent or improved transfection efficiencies and reproducibility over standard pre-plated methods. SureFECT is specifically optimized for reverse transfection in normal serum-containing medium, not serum-free medium like many other commercial transfection reagents. The simplicity of reverse transfection with the SureFECT Transfection Reagent allows you to quickly analyze gene function in your cell line of interest. The SureFECT Transfection Reagent may be used for reverse or traditional transfection of SABiosciences' SureSilencing shRNA Plasmids and Cignal™ Pathway Activity Reporters or for reverse transfection in SABiosciences' SureSilencing siRNA Arrays. Start optimizing the transfection efficiency of your cell line of interest with the SureFECT Transfection Reagent.

Need more information about SureFECT Transfection Reagent? please visit SureFECT Transfection Reagent Home or send Email to Technical Support

Kit Contents / Packing List / Storage Conditions

Please check the kit components immediately after you receive this package. SABiosciences is not responsible for any missing items not reported within two (2) business days upon receipt.

One tube of SureFECT Transfection Reagent

The SureSilencing SureFECT Transfection Reagent is shipped on ice.
For long-term storage, keep entire reagent at 4 ºC.

Shelf Life: This reagent is stable for 6 months after receipt if stored at the recommended temperature.

Brief Protocol
For experienced users ONLY. First time users should refer to the User Manual for the complete protocol.

Reverse Transfection (per well of a 96-well plate):

  1. Dissolve 100 to 200 ng nucleic acid in 96-well plates with 37 µL of Opti-MEM I per well by carefully rocking the plate back and forth several times.
  2. Dilute the SureFECT Transfection Reagent 1:10 in Opti-MEM I and mix.
  3. Add 3 µL to 6 µl of diluted transfection reagent to each well (corresponds to 3 µl undiluted SureFECT per g nucleic acid). Rock the plate back and forth, then left and right seven times each to thoroughly mix the nucleic acid and transfection reagent.
  4. Incubate the plate at room temperature for 20 minutes.
  5. Prepare your cells:
    a. For adherent cells, trypsinize and collect by centrifugation.
        For non-adherent cells, simply collect by centrifugation.
    b. Wash once with cell culture media by resuspension and re-centrifugation.
    c. Suspend in fresh cell culture media.
    d. Count the cells.
    Dilute to a concentration of 95 to 190 cells per µL with normal cell culture media.
    The final concentration may depend on your cell line and experimental conditions.
  6. When the incubation is finished, mix the cells very well.
    Add 160 µL of the cell suspension (15,000 to 30,000 cells ) to each well. Mix once.
    Shake each plate back and forth then left and right gently three times each.
  7. Incubate at 37C, 5% CO2 (or your cell line's normal growth conditions) for up to 48 or 72 hours. Note that some cell lines will require a media change 12 to 24 hours post-transfection.

Standard Transfection (per well of a 96-well plate):

  1. One day before transfection, seed 10000 to 15000 cells in each well of a 96-well plate (for roughly 30 to 40 percent confluence) with 100 µl to 150 µl of growth medium.
  2. On the day of transfection, add 0.2 µg of plasmid to 25 µl of Opti-MEM™ I Reduced-Serum Medium (Gibco). Mix gently and incubate at room temperature for 10 minutes.
  3. For each well, add 0.65 µl of SureFECT Transfection Reagent to 25 µl of Opti-MEM™ (i.e., 3.5 µl per µg plasmid).
    Mix gently and incubate at room temperature for 10 minutes.
  4. Add 25 µl of SureFECT Transfection Reagent in medium to each 25 µl plasmid in medium.
    Mix gently and incubate for 20 min at room temperature.
  5. Add each 50 µl volume to one well of cells growing in 100 µl to 150 µl of normal growth medium. Mix gently.
  6. Incubate the cells at 37 C in a CO2 incubator (or your cell line's normal growth conditions) for 24 to 48 hours. Note that some cell lines will require a media change 12 to 24 hours post-transfection.