Difference between revisions of "Pouring an agarose gel"
From Ucsbgalaxy
(Created page with "Use gloves! 50mL buffer ~0.5g agarose 1 uL ethidium bromide Mix agarose into buffer and heat until all agarose is incorporated. Wait until mixture is cool enough to hold comfor…") |
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1 uL ethidium bromide | 1 uL ethidium bromide | ||
| − | Mix agarose into buffer and heat until all agarose is incorporated. | + | # Mix agarose into buffer and heat until all agarose is incorporated. |
| − | Wait until mixture is cool enough to hold comfortably in hand then add ethidium bromide | + | # Wait until mixture is cool enough to hold comfortably in hand then add ethidium bromide |
| − | Rotate glass jar gently then cool | + | # Rotate glass jar gently then cool |
| − | Place rubber stoppers on the ends of the plastic channel | + | # Place rubber stoppers on the ends of the plastic channel |
| − | Insert comb to form wells, rounded edges up | + | # Insert comb to form wells, rounded edges up |
| − | Make sure the combs are at least 1mm away from the bottom | + | # Make sure the combs are at least 1mm away from the bottom |
| − | Pour agarose mixture into channel | + | # Pour agarose mixture into channel |
| − | Pour buffer into dish below the gel channel | + | # Pour buffer into dish below the gel channel |
| − | Wait until cool (gel will become cloudy) | + | # Wait until cool (gel will become cloudy) |
| − | Place Ladder into a well as a standard size (5uL) | + | # Place Ladder into a well as a standard size (5uL) |
| − | Mix loading buffer (1 uL) with the sample you are running (total ~5uL) on parafilm and load | + | # Mix loading buffer (1 uL) with the sample you are running (total ~5uL) on parafilm and # load |
| − | Turn meter on to ~100 Volts | + | # Turn meter on to ~100 Volts |
| − | Align channel so that the + side is farther from the samples | + | # Align channel so that the + side is farther from the samples |
| − | Let run until sample bands are about 1/2 way down the gel | + | # Let run until sample bands are about 1/2 way down the gel |
Latest revision as of 13:06, 15 November 2011
Use gloves!
50mL buffer ~0.5g agarose 1 uL ethidium bromide
- Mix agarose into buffer and heat until all agarose is incorporated.
- Wait until mixture is cool enough to hold comfortably in hand then add ethidium bromide
- Rotate glass jar gently then cool
- Place rubber stoppers on the ends of the plastic channel
- Insert comb to form wells, rounded edges up
- Make sure the combs are at least 1mm away from the bottom
- Pour agarose mixture into channel
- Pour buffer into dish below the gel channel
- Wait until cool (gel will become cloudy)
- Place Ladder into a well as a standard size (5uL)
- Mix loading buffer (1 uL) with the sample you are running (total ~5uL) on parafilm and # load
- Turn meter on to ~100 Volts
- Align channel so that the + side is farther from the samples
- Let run until sample bands are about 1/2 way down the gel
