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文献和实验Assembly of Nucleosomal Templates by Salt Dialysis
Because it is convenient to assemble nucleosomal templates through salt dialysis, large amounts of chromatin complexes can be made easily and in a short amount of time. This unit includes instructions for the various salt dialysis schemes (step versus gradient
setting) and collect the mononuclear cell (MNC) (buffy coat) layer. Resuspend and wash MNCs x 3 with Dulbecco's Phosphate Buffered Saline (DPBS) with 10% Fetal Bovine Serum (FBS) x 10 min, 515 g, before plating into two 12-well plates in full EPC growth
I have done. The protocol calls for 3 pig brains, and should yield ~ 60 mg of purified tubulin. Solutions and reagents: 100g P-11 cellulose phosphate fibrous cation exchanger (Whatman Inc, Clifton NJ) 6L 0.1M HCl 6L 0.1 M NaOH 2L 0.1 M MgSO4 2L 10
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