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文献和实验(Fisher M/4056/17)、Acetonitrile(Fisher A/0626/17)、Trypsin (Promega V528)、Trypsin resolve solution(Promega V530)、Ammonium bicarbonate(Sigma A6141)、ZipTip μC-18(Millipore ZTC18M096)、TFA(GE HealthCare)、0.2ml and 0.5ml EP tube(eppendorf)、50ml EP 0.5ml EP
, 10 μg/mL leupeptin, 1 mM phenylmethanesulfonylfluoride (PMSF), and 1 mM sodium orthovanadate. 6. Teflon cell scrapers (Fisher, Pittsburgh, PA ). 7. 2× SDS sample buffer for electrophoresis: 0.0375 M Tris-HCl, pH 6.5, 8% (w/v) SDS, 10% (v/v
Single and Double FISH protocols for Drosophila
, and then rinse 2-3 times with methanol. Embryos can be stored in methanol at -20oC for several months. 2.3 Post-Fixation and Hybridisation of Whole-mount Embryos The following steps are optimized for ~ 50 ul settled embryos in a 1.5 ml microfuge tube
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