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Team:METU Turkey/Results Discussion/Agarose Gel Electrophoresis
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Cerenseref (Talk | contribs) (New page: <br>MAIN STEPS/TIME TABLE <br>- 1% gel preparation [30 min] <br>- Running [1 h] <br>- Visualization [15 min] <br>MATERIALS <br>- Electrophoresis tank <br>- Power supply <br>- Transillumin...) |
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| Line 1: | Line 1: | ||
| - | <br>MAIN STEPS/TIME TABLE | + | <br><h3>MAIN STEPS/TIME TABLE</h3> |
<br>- 1% gel preparation [30 min] | <br>- 1% gel preparation [30 min] | ||
<br>- Running [1 h] | <br>- Running [1 h] | ||
<br>- Visualization [15 min] | <br>- Visualization [15 min] | ||
| - | <br>MATERIALS | + | <br><h3>MATERIALS</h3> |
<br>- Electrophoresis tank | <br>- Electrophoresis tank | ||
<br>- Power supply | <br>- Power supply | ||
| Line 20: | Line 20: | ||
| - | <br>SOLUTIONS | + | <br><h3>SOLUTIONS</h3> |
<br>TAE buffer | <br>TAE buffer | ||
<br>- Stock TAE electrophoresis buffer (50X) | <br>- Stock TAE electrophoresis buffer (50X) | ||
| Line 31: | Line 31: | ||
| - | <br>CHECK-LIST PROCEDURE | + | <br><h3>CHECK-LIST PROCEDURE</h3> |
<br>- Mix 2 uL DNA + 3 uL sterile dH2O + 1 uL loading dye on the parafilm | <br>- Mix 2 uL DNA + 3 uL sterile dH2O + 1 uL loading dye on the parafilm | ||
<br>- Load sample to the wells of 1% gel | <br>- Load sample to the wells of 1% gel | ||
| Line 39: | Line 39: | ||
<br>- After running of the samples record the gel image [go to SOPs-experimental for application of camera and record of image] | <br>- After running of the samples record the gel image [go to SOPs-experimental for application of camera and record of image] | ||
| - | <br>NOTES | + | <br><h3>NOTES</h3> |
<br>- Before adding of EtBr make sure the temperature of gel solution, its temperature should not be too high. | <br>- Before adding of EtBr make sure the temperature of gel solution, its temperature should not be too high. | ||
<br>- 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. | <br>- 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. | ||
| - | <br>TROUBLESHOOTING | + | <br><h3>TROUBLESHOOTING<h3> |
<br>Missing Bands | <br>Missing Bands | ||
Revision as of 03:44, 28 October 2010
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.
