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| RNA Techniques Forum Post and discuss RNA methods and RNA science related topics. RNA extraction, cDNA synthesis, RNA EMSA, and other RNA protocols. |
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#1
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| Hi guys, I had a problem using Ambion Megaclear for the first time to purify in-vitro transcribed RNA. I ran some of the RNA on a gel before purification and it was just fine. However, when I checked the RNA on a gel after using Megaclear, it came out as a giant smear meaning it was degraded sometime along the way. For those of you who have used this kit, do you know what I did wrong? |
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#2
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| Hello, That sounds like degradation. Be very careful you didn't use an old kit or a contaminated one. I would use Roche RNA sephadex G-25 or G-50 columns they are really good although do cost a bit (25-30 columns for $120?). They are stored in 4C and are RNAse fre. I have used them before for purification of in vitro transcribed RNA and it worked really well. |
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| megaclear , problem , purification , rna |
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