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25 lines
1.2 KiB
YAML
25 lines
1.2 KiB
YAML
name: 24-well Plate siRNA Transfection (RNAiMAX)
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id: PROT-0020
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version: 1.0
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description: Protocol for transfecting 4 siRNAs (including siNC) into cells in a 24-well plate using Lipofectamine RNAiMAX.
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author: Lab Agent
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created: 2024-06-08
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materials:
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- siRNA: siRNA#1 (final 10 nM)
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- siRNA: siRNA#2 (final 10 nM)
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- siRNA: siRNA#3 (final 10 nM)
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- siRNA: siNC (negative control, final 10 nM)
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- Lipofectamine RNAiMAX: 1.5 µL per well
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- Opti-MEM Reduced Serum Medium: as required
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- Cells: seeded in 24-well plate (e.g., 5x10^4 cells/well)
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steps:
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- "Seed cells in 24-well plate 24 hours prior to transfection (e.g., 5x10^4 cells/well)."
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- "For each siRNA, dilute 10 pmol siRNA in 50 µL Opti-MEM per well."
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- "In a separate tube, dilute 1.5 µL RNAiMAX in 50 µL Opti-MEM per well."
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- "Combine diluted siRNA and RNAiMAX, mix gently, and incubate for 10-20 min at room temperature."
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- "Add 100 µL siRNA-RNAiMAX complex dropwise to each well containing cells in 400 µL growth medium."
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- "Incubate cells at 37°C, 5% CO2 for 24-72 hours."
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notes: |
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- siNC is a non-targeting negative control siRNA.
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- Adjust cell number and reagent volumes as needed for specific cell lines.
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- Refer to manufacturer's instructions for further optimization. |