IGEM:Peking/2007/Switch: DNA electrophoresis
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 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.
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.
0.5M EDTA（pH 8.0）100mL ,0.05M EDTA（pH 8.0）
constant volume to 1L
1×TAE +EB 1L
0.04M Tris-AC+0.001M EDTA （pH 8.0）
10mg/mL EB 50uL 0.5ug/mL
10×Loading Buffer 50mL
20g 40% bromophenol blue
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 is a strong carcinogen.
In EB polluted area we should use gloves to touch anything.