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Agarose Gels
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==Agarose Gel== | ==Agarose Gel== | ||
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When making a small gel use 50mL of TBE buffer. The dispenser is on the bench behind ours in a large Nalgene container. Use the graduated cylinder next to it, but only for the TBE buffer. Pour that into a 100mL flask. Add the desired amount of agarose to (.4g for .8%, .5g for 10%, and 1g for 2%). Microwave the mixture for just under one minute with the weigh boat you used ontop of it so it doesn't bubble out. The agarose should be fully dissolved. If you ever mix it, make sure that you SLOWLY swirl it on the bench without picking it up. This will prevent bubbles. Wait till it has stopped steaming before adding the ethidium bromide into it. While you are waiting you can tape both up both sides of the gel tray with the comb in place. Once this is done you can slowly pour it into the gel tray. Let the gel sit (4C room is good) for ~40min before using. | When making a small gel use 50mL of TBE buffer. The dispenser is on the bench behind ours in a large Nalgene container. Use the graduated cylinder next to it, but only for the TBE buffer. Pour that into a 100mL flask. Add the desired amount of agarose to (.4g for .8%, .5g for 10%, and 1g for 2%). Microwave the mixture for just under one minute with the weigh boat you used ontop of it so it doesn't bubble out. The agarose should be fully dissolved. If you ever mix it, make sure that you SLOWLY swirl it on the bench without picking it up. This will prevent bubbles. Wait till it has stopped steaming before adding the ethidium bromide into it. While you are waiting you can tape both up both sides of the gel tray with the comb in place. Once this is done you can slowly pour it into the gel tray. Let the gel sit (4C room is good) for ~40min before using. | ||
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Revision as of 16:28, 15 July 2009
Agarose Gel
- 50mL 0.5x TBE buffer
- Agarose (To determine number of grams to use, multiply volume (50mL) by percentage of gel. For example, use 1.5g of agarose in a 3% agarose gel.)
- 2.5μL ethidium bromide
When making a small gel use 50mL of TBE buffer. The dispenser is on the bench behind ours in a large Nalgene container. Use the graduated cylinder next to it, but only for the TBE buffer. Pour that into a 100mL flask. Add the desired amount of agarose to (.4g for .8%, .5g for 10%, and 1g for 2%). Microwave the mixture for just under one minute with the weigh boat you used ontop of it so it doesn't bubble out. The agarose should be fully dissolved. If you ever mix it, make sure that you SLOWLY swirl it on the bench without picking it up. This will prevent bubbles. Wait till it has stopped steaming before adding the ethidium bromide into it. While you are waiting you can tape both up both sides of the gel tray with the comb in place. Once this is done you can slowly pour it into the gel tray. Let the gel sit (4C room is good) for ~40min before using.
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