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文献和实验Joel Huberman DNA isolation procedure with modifications by Bonny Brewer
to the bottom of the tube and suck up the liquid. It doesn't matter if a few beads get transferred. Rinse the beads twice with 1.5 volumes of fresh NIB. Pool and spin at 8 K in an SS-34 rotor for 20 min. 7. Resuspend pellet of nuclei, ghosts, and unbroken cells
Large-scale yeast transformation
. Resuspend yeast in the 90 mls LiAc/TE, put back in same flask (rinsed with sterile water). Shake at 30o C for 30 mins. Add 600 ul beta-mercaptoethanol. Shake at 30o C for 30 minutes. Add 1.7ml of 10mg/ml ss carrier DNA 3 . Cells ready
in a GSA rotor (5000 rpm, 5 min., RT). Discard the supernatant and resuspend the pellet in 10 ml T.E. Transfer the cells to a 50 ml centrifuge tube. Pellet the cells in an SS34 rotor (7000 rpm, 5 min., RT). Discard the supernatant and resuspend
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