I tried use a 2ml tissue grinder and optiprep density gradient, but didn't see the nuclei band. I wonder if the tissue grinder is too hard to deal with drg or the amount of drg I used is too low( I pooled 7 mouse DRG)
Bo Zhang Hi, I am currently trying to extract nuclei from ganglion (n=10). I am using the grinder to homogenize the tissue in lysis buffer. However, I hardly found nuclei in my final sample. I have the same confusion about whether my tissue lysis process was too harsh or whether it was the low number of ganglia. Were you able to solve your problem?
Gitalee Sarker Bo Zhang Did either of you ever solve this problem? I've been working on optimizing this in my lab, but I haven't been able to get more than ~180nuc/ul after homogenizing 14 DRGs in 500uL nuclear lysis buffer then adding 1000uL more lysis buffer before spinning it down. It feels like the numbers should be much higher given the number of cells in the DRG....I'm thinking my lysis might be too harsh as well.
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