Team:METU Turkey/Results Discussion/Agarose Gel Electrophoresis
From 2010.igem.org
Contents |
MAIN STEPS/TIME TABLE
- 1% gel preparation [30 min]
- Running [1 h]
- Visualization [15 min]
MATERIALS
- Electrophoresis tank
- Power supply
- Transilluminator
- Appropriate comb
- P10 and P100
- DNA Ladder 100-10 kb (Fermentas #SM0331)
- Agarose
- Loading dye
- TAE Buffer (1X)
- Distilled water [M14]
- Sterile dH2O [M14]
- EtBr
- Parafilm
SOLUTIONS
TAE buffer
- Stock TAE electrophoresis buffer (50X)
- Use 1X TAE
- 20 mL TAE
- 980 mL dH2O
1% Electrophoresis gel
- 0.5 gr Agarose
- 50 mL 1X TAE buffer (1%)
CHECK-LIST PROCEDURE
- Mix 2 uL DNA + 3 uL sterile dH2O + 1 uL loading dye on the parafilm
- Load sample to the wells of 1% gel
- Adjust the voltage of power supply to 75 V
- Adjust the time of power supply to 65 min
- Check transilluminator
- After running of the samples record the gel image [go to SOPs-experimental for application of camera and record of image]
NOTES
- Before adding of EtBr make sure the temperature of gel solution, its temperature should not be too high.
- Make sure the voltage is at the correct settings at all times and always use enough buffering solution to cover the gel in order to prevent inconsistent readings.