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Team:Newcastle/Gel electrophoresis
From 2010.igem.org
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==Procedure== | ==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 | + | # Make up 1% agarose gel (1 g of agarose in 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. | # 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. | # Transfer harden gel into the gel tank and add TAE buffer until the gel is completely submerged. | ||
| Line 25: | Line 27: | ||
# Loading buffer is then added together with the sample before loading onto the gel matrix. | # 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. | # Run gel at 90V until the desired separation is achieved and visualize using the gelDoc. | ||
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'''Go back to our [[Team:Newcastle/Protocol list|Protocol List]]''' | '''Go back to our [[Team:Newcastle/Protocol list|Protocol List]]''' | ||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Revision as of 08:19, 6 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 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, 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.
Go back to our Protocol List
   
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