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文献和实验Preparation of Brain Membrane Fractions
strokes). Never use a polytron.Centrifuge the homogenate at 800 x g for 20 min.Spin resulting supernatant at 16,000 x g for 30 min.Resuspend the pellet in 20 mM HEPES buffer (pH 7.4), rehomogenize and centrifuge at 16,000 x g for 15 min.Pool the pellets
Preparation of Brain Membrane Fractions for Western Blot Analysis
-FEP polymer homogenizer at 900 rpm (12�15 strokes). Never use a polytron. Centrifuge the homogenate at 800 x g for 20 min. Spin resulting supernatant at 16,000 x g for 30 min. Resuspend the pellet in 20 mM HEPES buffer
. We present all relevant methods including the initial site selection, molecular cloning, delivery of guide RNAs (gRNAs) and Cas9 into mammalian cells, verification of target cleavage, and assays for detecting genomic modification including indels
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