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| Good morning/evening/afternoon all! I have one simple question, i am working with RNA-dependent RNA polymerase, and in its assay was done as it should be, ive got a ladder of different RNA population, however i have a feeling that my RNA is stucked up in the wells.... afterwards i realized that my loading buffer does NOT contain formaldehyde that would react with -N= or :N- or any other possible target for nucleophilic attack, so the formamide work (denaturing the RNA at 65C 10 min) is totally useless! stucked up and i dont see a desired product (about 10KB)... Am I right? Another prob i have is that the resolution of my gel and its discreteness is much more to be desired, i use phosphate buffer, 6% formaldehyde, agarose system [BAD buffer goes hot and it stinks However, i tried to use MOPS... but it does not contain formaldehyde... got bad results, really bad.. I have heard about agarose-acrylamide gels, with a fantastic resolution? Are there?.. and one professor at our fascility says that it is a real art to make those gels, any comments on that? Could anyone please suggest me a very NICE approach to solve those problem? Thank You in advance, Drew |
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| agarose , formaldehyde , gel , rna |
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