DNA Transfection Reagent

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SuperFection™

DNA Transfection Reagent

INSTRUCTION MANUAL

Catalog No.

SL100566/SL100567

EASY PROTOCOL

Step 1:1x105 cells are seeded in 24-well plate in 360µl 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 0.5µg of DNA in 20µl of serum free DMEMmedium (or Opti-Medium from Invitrogen). Please avoid vortexing the DNA diluent. Then dilute SuperFection™1.5µl 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 complexdirectly to the cell growth medium. Incubate at 37°C and 5%CO2for 4 hours.

Step 4:Replace the DNA containing medium with fresh cellgrowth medium with 10%FBS and incubate at 37°C and 5%CO2for additional 24 hours or 48~72 hours as needed.

Step 5:Depending on the cell type and promoter activity. Theassay for the reporter gene can be performed 24~72 hoursfollowing 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.

QUICK REFERENCE

Table 1:Major Transfected Cell Types

HelaHEK293MDCK

HepG2NIH-3T3BHK-21

MA10CV1B16

COS-7CHOPC-12

MDA231COS-1AtT-20

Table 2:Volume of Transfection Reagents

DNA (µg) / DNA diluent DMEM (µl) / SuperFection(µl)
0.5 / 40 / 1.5
1 / 80 / 3
2 / 160 / 6
4 / 320 / 12
8 / 640 / 24

Table 3:Transfection Volume and siRNA Amount for Culture Dishes

Culture Dish Type / DNA (µg) / Transfection Volume (ml)
96-well / 0.1 ~0.4 / 0.15
24-well / 0.5 ~1.5 / 0.4
6-well / 2 ~5 / 1.2
60 mm / 5 ~8 / 3
100 mm / 8 ~12 / 6

Note:

1The data from above tables are for reference only.Actual amount can be adjusted foroptimization 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.

TEL. (301)-330-6381 FAX. (301)-560-4919 Email: