User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/07/05

AU CHEM-570 Lab Prep

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Objective Add today's objective(s) here... Bench work Analytical gel ladder -- Matt Hartings' sample - 6. -- 5μL 1:10 pTXB1His 2.5μL gel-purified double-digested insert from last week 2.5μL gel-purified double-digested vector(His) from last week 2.5μL gel-purified double-digested vector from last week 5μL V(His)+I ligation from last week 5μL V+I ligation from last week → 50' @ 90V → stain in EtBr Transformation 100μL DH10B + 5μL V(His)+I from last week (same as what is on gel) 100μL DH10B + 5μL V+I from last week (same as what is on gel) 100μL DH10B + 5μL V(His) neg ctrl from last week 100μL DH10B + 5μL V neg ctrl from last week heat shock is 45s @ 42°C plate 100μL on LBAmp100 → 37°C O/N Miniprep 10 mL LB + 10μL Amp100 + V+I in DH10B colony from last week's transformation (100μL plate) 10 mL LB + 10μL Amp100 + V+I in NovaBlue colony from last week's transformation, but the 450μL re-plate → 37°C O/N w/ shaking Results can't see undigested vector gel-purified samples did not float away and are visible on gel (good!) clearly see insert and double-inserts (~3kb). Faint band visible at correct size for vector+insert ligation (~7kb) but hard to see in photo.