**This is an old revision of the document!**
Protocol for RNA extraction using Trizol reagent
Based on the user guide of Trizol reagent by Invitrogen.
Material
- Sample : 6 x 10^6 of cell-line cells OR 20 mg of Tissue sample
| 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 with the kit.=
| 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 |
- Freshly prepared DNaseI solution included in the kit
| 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.