Based on the user guide of Trizol reagent by Invitrogen.
Material
Contents and storage
| Contents | Cat. No. 15596026 (100 reactions) | Cat. No. 15596018 (200 reactions) | Storage |
| TRIzolTM Reagent | 100 mL | 200 mL | 15–30°C |
Required items which is not supplied.
| Item | Memo |
| Equipment | |
| Water bath or heat block at 55–60°C | |
| Reagents | |
| Isopropanol | |
| Ethanol, 75% | |
| RNase-free water of 0.5% SDS | |
| (Optional) RNase-free glycogen | |
IMPORTANT!
Perform RNA isolation immediately after sample collection or quick-freeze samples immediately after collection and store at –80°C or in liquid nitrogen until RNA isolation.
| Sample type | Starting material per 1 mL of TRIzolTM Reagent | | |
| Tissues[1] | 50–100 mg of tissue | | |
| Cells grown in monolayer | 1 × 10 | 5 – 1 × 10 | 7 cells grown in monolayer in a 3.5–cm culture dish (10 cm2) |
| Cells grown in suspension | 5 – 10 × 10 | 6 cells from animal, plant, or yeasty origin or 1 × 10 | 7 cells of bacterial origin |
| | [1] Fresh tissues or tissues stored in RNAlaterTM Stabilization Solution (Cat. No. AM7020). | | |
| | | | |
| | | | |
| | | | |
| Check | Reagent | | 1 reaction (µL) | _ _ _ _ reaction (µL) | Memo |
| | ddH2O (RNAse free) | | 87.5 | | |
| | 10x DNase Buffer | | 10 | | |
| | DNaseI | | 2.5 | | 12 units / µL |
| | Total volume (µL) | | 100 | | _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ |
Protocol
1 Sample preparation
First step of sample preparation differs if you are using (A)culture cells OR (B)Tissue specimen.
Buffers are highlighted in blue to prevent mistakes.