Berkeley LBL/DNAGelElectrophoresis

From 2007.igem.org

< Berkeley LBL(Difference between revisions)
 
Line 2: Line 2:
242 g Tris Base
242 g Tris Base
 +
57.1 mL Glacial Acetic Acid
57.1 mL Glacial Acetic Acid
 +
600 mL ddH2O
600 mL ddH2O
 +
Mix. Bring volume to 1 L.  
Mix. Bring volume to 1 L.  

Latest revision as of 03:44, 27 October 2007

1. Prepare 50X TAE as:

242 g Tris Base

57.1 mL Glacial Acetic Acid

600 mL ddH2O

Mix. Bring volume to 1 L.

2. Make o.8% Agarose gel

3. Melt agarose in the microwave

4. Seal horizontal gel apparatus. Pour molten agarose onto gel plate to a depth of 4 - 8 mm.

5. Insert a comb until its base is 1 mm from the base of the gel. Allow to cool

6. Remove comb and submerge in 1X TAE buffer.

7. Prepare 6X Loading Dye as.

8. Add 1/5 volume 6X Loading Dye to sample. Mix well.

9. Add sample to well. Do not overflow well. For large samples, either reduce sample in volume prior to adding dye, or load into multiple wells.

10. Electrophorese until Bromophenol Blue is near the end of the gel. This dye runs at ~800 b.

11. Remove gel and visualize bands under uv light.