Give it a Chance
Never give up on a melted agarose gel. Just put it on ice 'til it solidifies and then take a look on the UV light. It just might be fine.
posted by Claire at 17:57
Friday, July 15, 2005
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- Name: Claire
Nature-loving, a capella-singing, high school-teaching, veterinary medicine-practicing member of generation Y
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2 Comments:
Absolutely. In fact, sometimes it takes just this sort of recrystallization to orient the aleoketone lattice. But (obviously) the degree of success depends upon the initial choice of TBE or TAE, the molarity of the agarose in the gel, and the maximum reheating temperature. Fortunately, one may always microverify the result by sending a UV-derived photonic sample through a diffuse reflectance spectrophotometer (with grating) and examining the peaks around 255 nm. If three or more peaks are clearly differentiated (i.e. with > 1 nm separation), you've got a problem. Otherwise [i.e. if you see no more than two, relatively large, peaks] you should regard the recrystallization as successful.
;-)
I hope no one thinks our heads are screwed on as straight as they will go when we are writing this kind of stuff...
You have spoken well, my brother, as you have many times in the past.
Always, always use TBE - its recrystallization characteristics are certainly unmatched. Optimization of the agarose molarity is of the utmost importance, although the exact molarity may be more or less impossible to obtain given the polymer nature of the compound. A high molarity leads to maximally melted gels due to the great resistance it provides against the current.
I don't know what we would do without UV photonic samples. They are highly feared as the causative agent of skin cancer, but the trusty light box and snazzy Bio Doc-It imaging system would be of no value without them. Spectrophotometers are great for measuring DNA concentrations in the final eluted samples following gel extraction. But for the initial recrystallization analysis, just use the ole' light box.
May the photons be with you.
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