Difference between revisions of "Dulbeccos Modified Eagle's Media, DMEM adding extras"
From mn.fysikk.laglivlab
(Created page with "== Dulbeccos Modified Eagle's Media, DMEM adding extras == * It is best to prepare a few tubes with the required amount of FETAL BOVINE SERUM (FBS) and Penicilin Streptomycin...") |
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| − | == | + | == Required equipment == |
| + | * 500 mL DMEM | ||
| + | * 50 mL FBS | ||
| + | * 5 mL Pen-Strep | ||
| + | * 0.2 µm filters | ||
| + | * 20 mL syringe | ||
| + | * 60 mL syringe | ||
| − | + | == Protocol == | |
| − | + | # Before prepare for DMEM solution, take out 1 tube of FBS and 1 tube pen-strep from the freezer and place them in the fridge (1 day in advance, after the overnight is fine). | |
| − | + | # Work inside the LAF bench and use syringes and 0.2 µm filter to add 50 ml FBS and 5 ml Pen-Strep to the 500ml DMEM. | |
| − | + | # Tighten the caps, and slightly shake the bottle to mix them. | |
| + | # Label the bottle, then store it in the fridge:<!-- | ||
| + | --><pre>name, dd/mm/yy, +10% FBS, +1% P/S </pre> | ||
| − | |||
| − | FETAL BOVINE SERUM (FBS) | + | |
| − | + | == Stock solution preparation == | |
| − | + | It is best to prepare a few tubes with the required amount of FETAL BOVINE SERUM (FBS) and Penicilin Streptomycin (Pen-Strep), and store them in the freezer. Each time only unfreeze the small tubes, instead of the whole bottle. As for 500 ml DMEM, it requires 50 mL (or 10%) of FBS and 5 mL (or 1%) of Penicilin Streptomycin (Pen-Strep). | |
| − | + | ||
| − | + | ||
| − | Penicilin Streptomycin (Pen-Strep) | + | '''FETAL BOVINE SERUM (FBS)''' |
| − | + | # Use 75% antibac ethanol alcohol to spray both hands with gloves, then the surface of the FBS bottle, the surface of 25 ml Serological Pipette, a few of 50 ml conical tube, and place them inside the LAF fume hood. | |
| + | # Inside the LAF hood, completely loose the conical tube caps and placed them in frame holder. Transfer 51 ml FBS into each conical tube, and tighten the caps. | ||
| + | # Label each conical tube:<!-- | ||
| + | --><pre> 50 mL FBS, dd/mm/yy, name </pre> | ||
| + | # Store tubes in the freezer. | ||
| + | |||
| + | |||
| + | '''Penicilin Streptomycin (Pen-Strep)''' | ||
| + | # Use 75% antibac ethanol alcohol to spray both hands with gloves, then the surface of the FBS bottle, the surface of 25 ml Serological Pipette, a few of 15 ml conical tube, and place them inside the LAF fume hood. | ||
| + | # Inside the LAF hood, completely loose the conical tube caps and placed them in frame holder. Transfer 5.5 ml Pen-Strep into each conical tube, and tighten the caps. | ||
| + | # Label each conical tube:<!-- | ||
| + | --><pre> 5 mL P/S, dd/mm/yy, name </pre> | ||
| + | # Store tubes in the freezer. | ||
Latest revision as of 12:34, 17 November 2023
Required equipment
- 500 mL DMEM
- 50 mL FBS
- 5 mL Pen-Strep
- 0.2 µm filters
- 20 mL syringe
- 60 mL syringe
Protocol
- Before prepare for DMEM solution, take out 1 tube of FBS and 1 tube pen-strep from the freezer and place them in the fridge (1 day in advance, after the overnight is fine).
- Work inside the LAF bench and use syringes and 0.2 µm filter to add 50 ml FBS and 5 ml Pen-Strep to the 500ml DMEM.
- Tighten the caps, and slightly shake the bottle to mix them.
- Label the bottle, then store it in the fridge:
name, dd/mm/yy, +10% FBS, +1% P/S
Stock solution preparation
It is best to prepare a few tubes with the required amount of FETAL BOVINE SERUM (FBS) and Penicilin Streptomycin (Pen-Strep), and store them in the freezer. Each time only unfreeze the small tubes, instead of the whole bottle. As for 500 ml DMEM, it requires 50 mL (or 10%) of FBS and 5 mL (or 1%) of Penicilin Streptomycin (Pen-Strep).
FETAL BOVINE SERUM (FBS)
- Use 75% antibac ethanol alcohol to spray both hands with gloves, then the surface of the FBS bottle, the surface of 25 ml Serological Pipette, a few of 50 ml conical tube, and place them inside the LAF fume hood.
- Inside the LAF hood, completely loose the conical tube caps and placed them in frame holder. Transfer 51 ml FBS into each conical tube, and tighten the caps.
- Label each conical tube:
50 mL FBS, dd/mm/yy, name
- Store tubes in the freezer.
Penicilin Streptomycin (Pen-Strep)
- Use 75% antibac ethanol alcohol to spray both hands with gloves, then the surface of the FBS bottle, the surface of 25 ml Serological Pipette, a few of 15 ml conical tube, and place them inside the LAF fume hood.
- Inside the LAF hood, completely loose the conical tube caps and placed them in frame holder. Transfer 5.5 ml Pen-Strep into each conical tube, and tighten the caps.
- Label each conical tube:
5 mL P/S, dd/mm/yy, name
- Store tubes in the freezer.
