Alm:Agarose gel electrophoresis: Difference between revisions
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(New page: Electrophoresis #Microwave TAE agarose 1% to melt. Pour 30 ml for a small gel into a small flask/beaker to cool a bit, then pour into the gel box with the correct comb for your size/numb...) |
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Electrophoresis | ==Overview== | ||
How to use gel electrophoresis to separate, measure and visualize DNA pieces. | |||
==Materials== | |||
*6X loading dye | |||
*[[TAE]] agarose 1% | |||
*1X TAE (~200ml) | |||
*gel box (casting tray optional) | |||
*power supply | |||
*[[Ethidium bromide]] staining solution | |||
*used TAE destaining solution | |||
*UV box/gel imager | |||
==Procedure== | |||
===Electrophoresis=== | |||
#Microwave TAE agarose 1% to melt. Pour 30 ml for a small gel into a small flask/beaker to cool a bit, then pour into the gel box with the correct comb for your size/number of samples. | #Microwave TAE agarose 1% to melt. Pour 30 ml for a small gel into a small flask/beaker to cool a bit, then pour into the gel box with the correct comb for your size/number of samples. | ||
| Line 7: | Line 23: | ||
#Slide on the lid and start the power supply. | #Slide on the lid and start the power supply. | ||
Staining and visualization | ===Staining and visualization=== | ||
*'''Wear blue/purple nitrile gloves when handling the items at the staining station.''' | |||
#Place the gel in the plastic box. | #Place the gel in the plastic box. | ||
#Stain for 1 hour in EtBr solution (concentration??). | #Stain for 1 hour in EtBr solution (concentration??). | ||
| Line 19: | Line 34: | ||
#Print out the image for your notebook, annotating the lanes, marker, and bands. | #Print out the image for your notebook, annotating the lanes, marker, and bands. | ||
#Note agreement with/variation from expected result. | #Note agreement with/variation from expected result. | ||
==Notes== | |||
==Contact== | |||
*Arne Materna | |||
*Sean Clarke | |||
==References== | |||
<!-- If this protocol has papers or books associated with it, list those references here. See the [[OpenWetWare:Biblio]] page for more information. --> | |||
<biblio> | |||
</biblio> | |||
[[Category:Protocol]] | |||
[[Category:In vitro]] | |||
[[Category:DNA]] | |||
[[Category:RNA]] | |||
[[Category:Protein]] | |||
[[Category:Alm]] | |||
Revision as of 13:48, 20 September 2007
Overview
How to use gel electrophoresis to separate, measure and visualize DNA pieces.
Materials
- 6X loading dye
- TAE agarose 1%
- 1X TAE (~200ml)
- gel box (casting tray optional)
- power supply
- Ethidium bromide staining solution
- used TAE destaining solution
- UV box/gel imager
Procedure
Electrophoresis
- Microwave TAE agarose 1% to melt. Pour 30 ml for a small gel into a small flask/beaker to cool a bit, then pour into the gel box with the correct comb for your size/number of samples.
- Add 6x loading dye to your samples.
- Make sure the red electrode is at the opposite end from the wells. You can check the progress of the DNA with the loading dyes to check the direc
- Set the time and settings on the power supply by pressing Mode to change fields. (typical settings for DNA: 90V constant voltage, 45 minutes)
- Slide on the lid and start the power supply.
Staining and visualization
- Wear blue/purple nitrile gloves when handling the items at the staining station.
- Place the gel in the plastic box.
- Stain for 1 hour in EtBr solution (concentration??).
- Pour the EtBr back into the light-proof bottle.
- Destain with used TAE for 2 minutes.
- Pour the TAE back into the bottle.
- Image the gel on a UV light box or the computerized gel imager in the Polz lab.
- Print out the image for your notebook, annotating the lanes, marker, and bands.
- Note agreement with/variation from expected result.
Notes
Contact
- Arne Materna
- Sean Clarke
