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Team:Newcastle/Gel electrophoresis
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| - | + | =Gel electrophoresis= | |
| + | [[Image:Newcastle Prep Chr Gel.jpg|250px|right]] | ||
==Materials== | ==Materials== | ||
| - | * | + | # 50X TAE buffer |
| + | #* Make up 1 liter of 50X TAE buffer with the following: | ||
| + | #* 242 g of TRIS base | ||
| + | #* 57.1 ml of acetic acid | ||
| + | #* 100 ml of 0.5 M of EDTA (pH 8.0) | ||
| + | #* Top up to 1 liter with water | ||
| + | # SafeView | ||
| + | # Agarose | ||
| + | # DNA ladder | ||
| + | # Eppendorf tube | ||
| + | # Gel making tank | ||
| + | # Gel running tank | ||
| - | == | + | ==Procedure== |
| - | # Make up 1% agarose gel (1 g of agarose | + | # Make up 1% agarose gel (1 g of agarose per 100 ml of TAE buffer) and using the microwave to dissolve the agarose. |
| - | # Wait for 30 min to allow the gel to harden | + | # Allow the molten agarose to cool down before adding 5 μl of Safeview dye. |
| - | # Transfer harden gel into the gel tank and add TAE buffer until the gel is completely | + | # Set up the gel tray containing the gel comb and transfer 60 ml of molten agarose gel into the gel tray (take care not to cover the gel comb). |
| - | # Depending on the nature of the sample, | + | # Wait for 30 min to allow the gel to harden. |
| - | # Loading buffer | + | # Transfer harden gel into the gel tank and add TAE buffer until the gel is completely submerged. |
| - | # Run gel at 90V until separation is achieved and visualize using the gelDoc | + | # Depending on the nature of the sample, 3 μl of GeneRuler™ 1 kb Plus DNA Ladder is used for analysing DNA. |
| + | # Loading buffer is then added together with the sample before loading onto the gel matrix. | ||
| + | # Run gel at 90V until the desired separation is achieved and visualize using the gelDoc. | ||
| + | |||
| + | |||
| + | '''Go back to our [[Team:Newcastle/Protocol list|Protocol List]]''' | ||
| + | |||
| + | {{Team:Newcastle/footer}} | ||
Latest revision as of 14:28, 26 October 2010
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Gel electrophoresis
Materials
- 50X TAE buffer
- Make up 1 liter of 50X TAE buffer with the following:
- 242 g of TRIS base
- 57.1 ml of acetic acid
- 100 ml of 0.5 M of EDTA (pH 8.0)
- Top up to 1 liter with water
- SafeView
- Agarose
- DNA ladder
- Eppendorf tube
- Gel making tank
- Gel running tank
Procedure
- Make up 1% agarose gel (1 g of agarose per 100 ml of TAE buffer) and using the microwave to dissolve the agarose.
- Allow the molten agarose to cool down before adding 5 μl of Safeview dye.
- Set up the gel tray containing the gel comb and transfer 60 ml of molten agarose gel into the gel tray (take care not to cover the gel comb).
- Wait for 30 min to allow the gel to harden.
- Transfer harden gel into the gel tank and add TAE buffer until the gel is completely submerged.
- Depending on the nature of the sample, 3 μl of GeneRuler™ 1 kb Plus DNA Ladder is used for analysing DNA.
- Loading buffer is then added together with the sample before loading onto the gel matrix.
- Run gel at 90V until the desired separation is achieved and visualize using the gelDoc.
Go back to our Protocol List
   
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