Comb size
- Single wells from the smallest comb (15 wells per row) can hold up to 20 ul
- Single wells from un-taped medium comb (8 wells per row) can hold up to 50 ul
Agarose density
- Above 3 kb: 0.8%
- 1-3 kb: 1%
- 250-1000 bp: 1.5%
- Below 250 bp: 2%
- 75V for 45 min, or 85V for 35 min
- Minimizing the amount of agarose in the cutout band seems to improve yields
Notes
- I clean the razor blade between each use with a Kimwipe and do not change blades until it is dull or known to be contaminated. I have never had cross-contamination in my gel extraction doing it this way. --YC
- Using the nano-drop to determine DNA concentration from a gel extraction is inaccurate. If you really need a concentration, run a small sample of the eluent on a gel and determine the concentration by comparing the brightness to that of the ladder at known concentration.
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