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文献和实验Constructing a Low-budget Laser Axotomy System to Study Axon Regeneration in C. elegans
to control focus 1) This system uses dual Galilean beam expanders to expand the laser beam to fill the back aperture of the objective (10 mm). Mount the 4 lenses on to carriers and attach them to the rail (L1f1 L2f2 and L3f1' L4f2'). Adjust
buffer formula) 10 X REact buffer 2 : for, e.g. Hind III and Sst 1 (5 ml) 1 X Conc. Stock 500 mM 1 M Tris (pH 8.0) 2.5 ml 100 mM 1 M MgCl2 0.5 ml 500 mM NaCl 0.5 ml 25 mM Spermidine(3HCl) 0.032 g H2O 1.5 ml Mix, aliquote 2 ml
µL 5 M K-acetate, vortex, incubate on ice for 20 min. Spin @ 12K x g in microfuge for 10 min. 4) Transfer 750-800 µL supernatant to new microfuge tube and add an equal vol of isopropanol. Immediately spin down nucleic acids by centrifugation
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