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Topic: DNA Ligase reaction  (Read 4295 times)

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Offline lily_m

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DNA Ligase reaction
« on: October 21, 2009, 03:29:02 PM »
Hello all, can someone please help me with this question. I conducted a DNA ligase experiment with 1 μl of digested pUC18 DNA with 1 μl of DNA ligase enzyme and ran the ligated sample next to a control with agarose gel electrophoresis. My problem is that the ligated sample has appeared to decrease in band size in comparison to the control. I expected an increased due to the joining of DNA fragments. Can anybody please tell me why this is so??

Offline Yggdrasil

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Re: DNA Ligase reaction
« Reply #1 on: October 21, 2009, 05:53:40 PM »
Are you converting a linear DNA into a circular DNA?  Circular DNAs tend to run faster than linear DNAs on agarose gels and could account for the higher mobility of your ligated sample.

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