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Southern Blot of a large DNA fragment using DIG

Southern blot

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#1 LrrOfOmicronPersei8

LrrOfOmicronPersei8

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Posted 21 September 2012 - 12:22 PM

Hello,
I'm having trouble with a southern blot on fragments larger than 10kb using DIG labeled probes. I think the problem is with the transfer. I am depurinating with 0.25M HCl for 20 min. Has anyone ever tried UV irradiation to fragment the DNA? Would transfering longer (say over 2 nights) be better or worse? Does anyone have experience with this?
Thanks in advance.





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