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| Agarose Gel Electrophoresis Forum Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels. |
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#1
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| Recently I came across a problem when preparing the agarose gel. Specifics: After boiling the agarose mixture (agarose + TBE) via microwave, I observed that there were many microbubbles in the gel. I decided to do the gel anyway, the end result was poor migration of DNA bands, specifically that parts of the DNA bands seem to lag behind considerably and looked similar to using a toothpick to drag a blotch of spilled ink around. Any ideas? (I will upload a picture as soon as I get to work) |
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#2
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| normally after pouring the heated agarose mixture onto gel cast, u can use a pipette tip to push microbubbles to corner of the gel. this can prevent bubbles obstructing the migration of you DNA. good luck. |
| The Following User Says Thank You to JennYeap For This Useful Post: | ||
admin (08-14-2011)
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#3
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| dear i suggest you to reduce the boiling time in microwave if you are heating the agarose for 2 min. in microwave reduce the time to one minute if solution is not clear then additional half min. should be given n also u can remove bubbles with pipette tips |
| The Following User Says Thank You to SAFIA For This Useful Post: | ||
admin (08-14-2011)
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#4
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| thanks for the replies guys. It turns out that the agarose was not completely dissolved in the solution. |
| The Following User Says Thank You to oblongata For This Useful Post: | ||
admin (08-14-2011)
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#5
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| Great tips above, I would not over heat it as well - take breaks during microwaving as it can get super hot and bubble |
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#6
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| 1- dont let it cool below 50C- 2- then add Et. Bromide and swirl the flask on the Bench |
| Tags |
| agarose , bubbles , gel , reducing , setting |
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