| 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 SuperArray's SureSilencing shRNA Plasmids and Cignal™
Pathway Activity Reporters or for reverse transfection in SuperArray's
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. SuperArray 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):
- 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.
- Dilute the SureFECT Transfection Reagent 1:10 in Opti-MEM I and mix.
- Add 3 µL to 6 µl of diluted transfection reagent to each well
(corresponds to 3 µl undiluted SureFECT per µg nucleic acid). Rock the
plate back and forth, then left and right seven times each to thoroughly mix
the nucleic acid and transfection reagent.
- Incubate the plate at room temperature for 20 minutes.
- 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.
- 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.
- Incubate at 37°C, 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):
- 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.
- 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.
- 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.
- 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.
- Add each 50 µl volume to one well of cells growing in 100 µl to 150 µl of
normal growth medium. Mix gently.
- 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.
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