IGEM:Peking/2007/Switch: DNA electrophoresis

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DNA eletrophoresis

0.5cm slot, 2ng is the bottom line for testing.


Lambda DNA/EcoRI+HindIII Marker Fermentas


Transgen 100bp DNA Ladder


Transgen 1kb DNA Ladder


Agarose concentrations

agarose concentration[%(w/v)]: linear DNA effective distinguishment[kb]

0.3  : 5-60

0.6  : 1-20

0.7  : 0.8-10

0.9  : 0.5-7

1.2  : 0.4-6

1.5  : 0.2-3

2.0  : 0.1-2


distinguish different size of DNA. EB moves opposite direction to DNA and will run out of the gel if the time for electrophoresis is too long. Under such situation, the gel should put in 1×TAE+EB solution for 30min before imaging.

Loading Buffer

In 0.5-1.4% agarose,the speed of bromophenol blue is the same as 300bp double strand linear DNA,the speed of Xylene blue is the same as 4kb double strand linear DNA.


50×TAE 1L

Tris 242g

2M Tris-AC

HAC 57.1mL

0.5M EDTA(pH 8.0)100mL ,0.05M EDTA(pH 8.0)

constant volume to 1L

1×TAE +EB 1L

50×TAE 20mL

0.04M Tris-AC+0.001M EDTA (pH 8.0)

10mg/mL EB 50uL 0.5ug/mL

H2O 980mL

10×Loading Buffer 50mL

20g 40% bromophenol blue

glycerol 25mL

constant volume to 50mL

method of making gel

Use 1×TAE+EB to make the gel, 1×TAE as the eletrophoresis buffer

The agarose should be shaken up and pour out as the gel

the slot lies at the negative electrode

the voltage should be remained constant during electrophoresis

EB pollution

EB is a strong carcinogen.

In EB polluted area we should use gloves to touch anything.