"
Team:Newcastle/Gel electrophoresis
From 2010.igem.org
(Difference between revisions)
Swoodhouse (Talk | contribs) (→Gel electrophoresis) |
(→Materials) |
||
| (10 intermediate revisions not shown) | |||
| Line 2: | Line 2: | ||
=Gel electrophoresis= | =Gel electrophoresis= | ||
| - | [[Image:Newcastle Prep Chr Gel.jpg| | + | [[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== | ==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. |
| + | # 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 | + | # Transfer harden gel into the gel tank and add TAE buffer until the gel is completely submerged. |
| - | # Depending on the nature of the sample, | + | # Depending on the nature of the sample, 3 μl of GeneRuler™ 1 kb 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. |
Latest revision as of 14:28, 26 October 2010
| |||||||||||||
| |||||||||||||
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
   
|
    
|
