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  2. → Viewing Profile: Topics: LrrOfOmicronPersei8

LrrOfOmicronPersei8

Member Since 27 Aug 2012
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Topics I've Started

Southern Blot of a large DNA fragment using DIG

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.

my human iPS cells have a lot of differentiation

27 August 2012 - 04:54 PM

My iPS cells keep differentiating spontaneously.  I'm on mTeSR/Matrigel and I am able to maintain alot of undifferentiated iPS cells but I always get quite a bit of differentiation.  I haven't had too much experience so how much differentiation is normal?  What is the best way to passage these?  For me the best way is using dispase, manually picking the good colonies, and breaking them down by gently pipetting.  But is there a better way?  Has anyone tried Invitrogen's EZpassage tool?
Thanks in advance

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