Team:DTU-Denmark/Gel preparation and gel electrophoresis
From 2011.igem.org
(Difference between revisions)
(Created page with "== Gel preparation (1% gel) (100 ml of the buffer) == # 100 ml of 1x TBE buffer . # 10 μl of ethidium bomide (10 mg/ml). # 1 g of agarose. # Assembled gel container. # Mix buf...") |
|||
Line 1: | Line 1: | ||
== Gel preparation (1% gel) (100 ml of the buffer) == | == Gel preparation (1% gel) (100 ml of the buffer) == | ||
- | + | <ol> | |
- | + | <li value="a">100 ml of 1x TBE buffer.</li> | |
- | + | <li value="b">10 μl of ethidium bomide (10 mg/ml).</li> | |
- | + | <li value="c">1 g of agarose.</li> | |
+ | <li value="d">Assembled gel container.</li> | ||
+ | </ol> | ||
# Mix buffer with agarose and heat in microwave until the solution is clear | # Mix buffer with agarose and heat in microwave until the solution is clear | ||
Line 10: | Line 12: | ||
# Place gel container in the electrophoresis machine, remove combs and cover with the TBE buffer. | # Place gel container in the electrophoresis machine, remove combs and cover with the TBE buffer. | ||
Gel electrophoresis: | Gel electrophoresis: | ||
- | + | <ol> | |
- | + | <li value="i">2μl of DNA sample.</li> | |
- | + | <li value="ii">3 μl of distilled water.</li> | |
- | + | <li value="iii">1μl of 6x loading dye.</li> | |
+ | <li value="iv">4.2 μl of Gene Ruler DNA ladder mix from Fermentas.</li> | ||
+ | </ol> | ||
+ | |||
It is recommended to use 7V for each cm of the gel length and to run the gel for 45 min. | It is recommended to use 7V for each cm of the gel length and to run the gel for 45 min. |
Revision as of 14:10, 18 September 2011
Gel preparation (1% gel) (100 ml of the buffer)
- 100 ml of 1x TBE buffer.
- 10 μl of ethidium bomide (10 mg/ml).
- 1 g of agarose.
- Assembled gel container.
- Mix buffer with agarose and heat in microwave until the solution is clear
- Add 10μl of ethidium bromide (10mg/ml).
- Pour solution to the gel container and leave it to solidify (30-45 min).
- Place gel container in the electrophoresis machine, remove combs and cover with the TBE buffer.
Gel electrophoresis:
- 2μl of DNA sample.
- 3 μl of distilled water.
- 1μl of 6x loading dye.
- 4.2 μl of Gene Ruler DNA ladder mix from Fermentas.
It is recommended to use 7V for each cm of the gel length and to run the gel for 45 min.