I'm digesting a few different mammalian tissues with a spin kit and proteinase k. Some tissues (esp. testis and pituitary gland) look fully digested and clear. But once the lysis buffer is added, the "glob" appears. We have to remove it, or else it interferes with the spin column. What is this glob? We haven't had DNA yield issues, except with testis tissue (same starting tissue amount). Is the tissue not being fully digested, or is it some type of lipid? Might it have something to do with the testis DNA yield?
Have you tried to homogenize your tissue before extraction? Prot K is not able to fully digest a (even small) chunk of tissue efficiently. During the digestion you may need to shake your tube occasionally.
I concur
You can also digest at 55c for 1 hour or 37C over night with no deleterious affect
If your insoluble matter remains after homogenisation and long digestion with pulse spins and inverting tubes every few min, simply separate clear spt and column purify
For large tissue volumes you could even try trizol extraction before column purification
But as mentioned i would start with homogenisation
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