RNAi專用的轉染試劑
RNAifectin™ Transfection Reagent, G073
【萊恩生物科技有限公司04-25239639】abmgood台灣總代理
| Cat.No.: | G073 |
| Quantity: | 1.0ml (100-500 transfections) |
Description
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Escpecially developed for the efficient transfection of eukaryotic cells with RNAi oligo's. This transfection reagent is perfect for any of your RNAi transfection needs.
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Application
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RNAi knockdown experiments in a wide variety of cell lines (HeLa, HEK-293 etc). Highly specific and non-toxic transfection.
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Protocol
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Storage
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Store at 4ºC. Do not freeze.
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Notes
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This product is distributed for laboratory research only. Caution: Not for diagnostic use .
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產品介紹 Description
ANAifectin™ is a transfection reagent specially formulated with multiple cationic polymers. It is suitable for the transfection of RNAi oligoes into cultured eukaryotic cells.
RNAi轉染操件手冊:
These conditions are recommended as guidelinesonly. A 6-well or 35mm dish is adequate for most applications, but larger vessels are sometimes required. In that case, consult table 1.
1. In a 6-well plate, seed cells at a density of 1-3x10⁵ per well in 2.0ml of the appropriate
growth medium (with serum if cells are cultured in presence of serum). Incubate the cells at 37°C until cells are 60-90% confluent. This will usually take 18-24 hours, depending on cell types. Since transfection efficiency is sensitive to culture confluence, it is important to maintain a standard seeding protocol from experiment to experiment
2. Prepare the following solutions in sterile 12x75 mm or microcentrifuge tubes: Solution A: For each transfection, dilute 1-3μg of RNAi oligoes into 125μl serum-free
medium. Solution B: For each transfection, dilute 4-10μl of
RNAifectin™ reagent into 125μl of serum-free medium. (Vortex RNAifectin™ reagent before use)
3. Combine the solutions, mix thoroughly and incubate at room temperature for 20 mins to
allow DNA-lipsome complexes to form.
4. Remove growth medium from the cells and add 800μl of serum-free medium to the cells.
5. Add the RNAifectin™-oligo complexes to thecells, mix gently to ensure uniform distribution
and incubate at 37°C for 2-24 hours. We recommend starting with 5 hours.
6. Remove the transfection solution and add 2.0ml of the appropriate complete growth medium(with serum) to each well. Incubate cells at 37°Cfor a total of 24-72 hours.
7. Assay cell extracts for gene activity 24-72 hours aer the start of transfection depending on the cell types and promoter activity.
8. A similar procedure can be used to transfect an RNAi vector for stable expression. At 72 hours after transfection, split the cells at a ratio of 1:10 into the selective medium for the marker gene transfected.
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