Cat # SL Store at 4 0 C GenJet™ siRNA Transfection Reagent Small 0.5 ml Large 1.0 ml Gaither Drive Gaithersburg, MD FAX TEL Toll Free Web: Description: GenJet™ siRNA Transfection Reagent, a hybrid of liposome and polymer, ensures superior efficiency siRNA delivery with invisible cytotoxicity. Based on innovative and proprietary lipid-conjugation and polymer synthesis technology and formulation, this product exhibits significant difference from other siRNA transfection reagents in the market. Quick Protocol: Step 1: 1x10 5 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 0 C and 5 % CO 2. The plate should be 60~70% confluent on the day of transfection. One hour before transfection, the serum-containing medium is replaced with 360 µl serum-free DMEM Medium with High Glucose. Step 2: For each well of 24-well plate, dilute 0.2 µg of siRNA in 40 µl of serum-free DMEM Medium with High Glucose. Vortex for 2 seconds and briefly spin down. Then dilute GenJet™ Reagent 2.0 µl to 40 µl serum-free DMEM Medium with High Glucose. Vortex for 2 seconds and briefly spin down. Drop the diluted GenJet™ reagent into diluted siRNA solution immediately. Important ! do not mix the solutions in the reverse order ! Step 3: Vortex the two solutions immediately and spin down briefly to bring drops to the bottom of the tube. Incubate for 10 minutes at room temperature. Important ! do not leave the mixture longer than 15 minutes ! Step 4: Add the mixture of GenJet™/siRNA complex directly to the serum-free DMEM Medium with High Glucose. Incubate at 37 0 C and 5% CO 2 for 4 hours. Step 5: Replace the siRNA containing medium with fresh cell growth medium with 10 % FBS and incubate at 37 0 C and 5 % CO 2 for additional 24 hours or 48~72 hours as needed. Step 6: Most RNA interference can be detected within 24 hours following transfection. Note: 1. The above transfection protocol is for 24-well plate. Other dish types refer to Table 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/GenJet ™ ratio. This product is for laboratory research ONLY and not for diagnostic use  2006 SignaGen Laboratories 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 Table 3: Transfection Volume and siRNA Amount for Culture Dishes Note: 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. Storage: Upon arrival store this product at 4 0 C. If stored properly, the product is stable for 18 months or longer. Product shipped at ambient temperature. siRNA (µg)siRNA Diluent DMEM (µl) GenJet™ Reagent (µl) Culture Dish TypesiRNA (µg)Transfection Volume (ml) 96-well well0.15 – well0.8 – mm1.5 – mm3.0 –