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Team:Newcastle/Gel electrophoresis
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# Make up 1% agarose gel (1 g of agarose in 100 ml of TAE buffer) and transfer 60 ml of molten agarose gel into the gel tray with 3 μl of Safeview dye. | # Make up 1% agarose gel (1 g of agarose in 100 ml of TAE buffer) and transfer 60 ml of molten agarose gel into the gel tray with 3 μl of Safeview dye. | ||
# Wait for 30 min to allow the gel to harden. | # 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 | + | # Transfer harden gel into the gel tank and add TAE buffer until the gel is completely submerged. |
| - | # Depending on the nature of the sample, 3μl of GeneRuler™ 1kb Plus DNA Ladder | + | # Depending on the nature of the sample, 3μl of GeneRuler™ 1kb Plus DNA Ladder is used for analysing DNA. |
| - | # Loading buffer | + | # Loading buffer is then added together with the sample before loading onto the gel matrix. |
| - | # Run gel at 90V until separation is achieved and visualize using the gelDoc. | + | # Run gel at 90V until the desired separation is achieved and visualize using the gelDoc. |
Revision as of 12:14, 5 August 2010
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Gel electrophoresis
Materials
- 50X TAE buffer
- Make up 1 liter of 50X TAE buffer with the following:
- 242g 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
- Gel making tank
- Gel running tank
Procedure
- Make up 1% agarose gel (1 g of agarose in 100 ml of TAE buffer) and transfer 60 ml of molten agarose gel into the gel tray with 3 μl of Safeview dye.
- 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, 3μl of GeneRuler™ 1kb 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.
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