Transfection Reagent GenJet™ In vitro siRNA Small 0.5 ml Large 1.0 ml 15875 Gaither Drive Gaithersburg, MD 20877 FAX. 301-560-4919 TEL. 301-330-5966 Toll Free. 1-866-918-6812 Email: info@signagenlabs.com Web: www.signagenlabs.com Cat # SL100570 Store at 4 0C This product is for laboratory research ONLY and not for diagnostic use Description: GenJet™ siRNA Transfection Reagent is liposome based transfection regent which ensures superior efficiency siRNA delivery with invisible cytotoxicity. Based on innovative and proprietary lipid-conjugation technology and formulation, this product exhibits significant difference from other siRNA transfection reagents in the market. Quick 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 0C and 5 % CO2. The plate should be 30~50% confluent on the day of transfection. Step 2: For each well of 24-well plate, dilute 8.8 ng of siRNA in 40 µl of serum-free DMEM Medium with High Glucose. Gently tap to mix. Please avoid vortexing the siRNA diluent. Then add GenJet™ Reagent 2.0 µl to the 40 µl diluted siRNA solution. Pipette up-down the mixture for 4~5 times, and incubate the mixture at room temperature for 15 minutes to allow siRNA complex generation. Note: For optimal formation of GenJet™/siRNA complex, use serum-free DMEM Medium with High Glucose to dilute siRNA and GenJet™ reagent! Step 3: Add the mixture of GenJet™/siRNA complex directly to the cell growth medium. Incubate at 37 C and 5% CO2 for 5 hours. Step 4: Replace the siRNA containing medium with fresh cell growth medium with 10 % FBS and incubate at 37 0C and 5 % CO2 for additional 24 hours or 48~72 hours as needed. Step 5: Most RNA interference can be detected within 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/GenJet™ ratio. 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 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 0C. If stored properly, the product is stable for 12 months or longer. Product shipped at ambient temperature. siRNA (ng) siRNA Diluent DMEM (µl) GenJet™ Reagent (µl) 8.8 40 2 22 80 5 46 160 10 98 320 20 200 640 Culture Dish Type siRNA Per Well (ng) Transfection Volume (ml) 96-well 2.6 0.2 24-well 8.8 0.4 6-well 31 1.2 60 mm 62 3.0 100 mm 147 6.0  2007 SignaGen Laboratories