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文献和实验dishes (e.g., Corning® CellSTACK®). 6. CO 2 incubator and biosafety cabinet. 7. Filter bottle, 0.5–1 L, 0.2-μm pore size (Corning or equivalent). 2.2. Preparation of Wnt3A CM for Fractionation 1. 20% (v/v) Triton X-100. 2. 1 M Tris-HCl, pH
Northern Blotting (Howell Lab)
before quantitation. Pipet up and down several times before taking 2 µl into 998 µl water. Read at 260 nm. RNA concentration (µg/µl) = OD x 500 x 0.04 µg/µl = OD x 20 Formaldehyde gel Boil 2 g agarose in 144 ml water (check weight
Northern Blotting方法(Howell Lab)
on ice for 5 min. e) Add 2 µl loading buffer and 1 µl 0.4 mg/ml EtBr. For the ladder lanes, use 3 µg of 1 kb ladder and treat the same way as with the samples. Running the gel For the Hoefer, run between 70-85 V for about 3 hr
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