b*********t
发帖数: 877 | 1
depending on the sizes of the bands (usually two). if they are both big and
close in size, then you will have to run it really slow for a long time (i
have one 11kb/9kb southern that i run at 40V for 10 hrs!); otherwise, you
can run it much faster (70V, 3 hrs.)
this should have been determined BEFORE you make your transgenic mice.
you should be able to predict the sizes/conditions based on your original
constructs.
depending on your probe. usually two bands: one endogenous, one transgenic.
good | M****e
发帖数: 70 | 2 refer to NAR 1991 19:42 P.W.Laird et al
i have tried to optimize the condition and it works pretty
well for me. basically, cut ~5mm tail when the mickeys are
about 14-d-old (do not forget ear tag), if you worry about
cross-contamination, clean scissors with 70% ethanol and
flame, otherwise, i typically clean it after collecting 5
samples. digest the tail in a tightly sealed epp. with 500ul
lysis buffer, 5ul proteinase K (10mg/ml). rotate O/N at 55C,
though 6hrs is pretty much sufficient. spin 10 |
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