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SuperFection™
DNA Transfection Reagent
QUICK REFERENCE
Table 1: Major Transfected Cell Types
Hela
HEK293
MDCK
HepG2
NIH-3T3
BHK-21
MA10
CV1
B16
COS-7
CHO
PC-12
MDA231
COS-1
AtT-20
INSTRUCTION MANUAL
Catalog No.
SL100566/SL100567
Table 2: Volume of Transfection Reagents
EASY PROTOCOL
Step 1: 1x105 cells are seeded in 24-well plate in 360
µl of appropriate growth medium containing serum and
antibiotics on the day before transfection. Incubate
the cells at 37 °C and 5% CO2. The plate should be
60~80% confluent on the day of transfection. One
hour before transfection, the serum-containing medium
is replaced with 360 µl Opti-Medium (Invitrogen) or
DMEM serum-free medium.
Step 2: For each well of 24-well plate, dilute 0.5 µg of
DNA in 20 µl of serum free DMEM medium (or OptiMedium from Invitrogen). Please avoid vortexing the
DNA diluent. Then dilute SuperFection™ 1.5 µl to 20 µl
Opti-Medium and gently tap the tube to mix. 5
minutes later, mix the diluted DNA and SuperFection™
reagent followed by 20 minutes incubation at room
temperature to allow DNA complex generation.
Step 3: Add the mixture of SuperFection™/DNA
complex directly to the cell growth medium. Incubate
at 37°C and 5% CO2 for 4 hours.
Step 4: Replace the DNA containing medium with
fresh cell growth medium with 10% FBS and incubate
at 37 °C and 5 % CO2 for additional 24 hours or
48~72 hours as needed.
Step 5: Depending on the cell type and promoter
activity.
The assay for the reporter gene can be
performed 24~72 hours following transfection.
Note:
1. The above transfection protocol is for 24-well plate.
Other dish types refer to Table 3.
2. The protocol is optimized for adherent cell lines
tested. To achieve the highest efficiency for specific
cell(s), more optimization may be necessary.
3. The major factors for transfection optimization
include siRNA quantity and siRNA/SuperFection ratio.
www.signagenlabs.com
TEL. (301)-330-6381
DNA
(µg)
0.5
1
2
4
8
DNA diluent DMEM
(µl)
40
80
160
320
640
SuperFection(µl)
1.5
3
6
12
24
Table 3: Transfection Volume and siRNA Amount
for Culture Dishes
Culture Dish
Type
96-well
24-well
6-well
60 mm
100 mm
DNA (µg)
0.1 ~0.4
0.5 ~1.5
2 ~5
5 ~8
8 ~12
Transfection
Volume (ml)
0.15
0.4
1.2
3
6
Note:
1 The data from above tables are for reference only.
Actual amount can be adjusted for optimization
according to experimental conditions.
LIMITED WARRANTY
NO OTHER WARRANTIES OF ANY KIND, EXPRESS OR IMPLIED,
INCLUDING WITHOUT LIMITATION, IMPLIED WARRANTIES OF
MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE
PROVIDED BY SIGNAGEN. SIGNAGEN SHALL HAVE NO LIABILITY
FOR ANY DIRECT, INDIRECT, CONSEQUENTIAL OR INCIDENTAL
DAMAGES ARISING OUT OF THE USE, THE RESULTS OF USE, OR
INABILITY TO USE THIS PRODUCT.
This product is for research ONLY and must be used
according to the related regulations.
PACKAGE 0.5 ml/1.0 ml per vial. It contains sufficient reagent for
330/660 reactions based on transfecting 500 ng of RNA in 24-well
plate.
STORAGE Store at 4 °C. If stored properly, the product is stable
for 12 months or longer.
FAX. (301)-560-4919
Email: [email protected]