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100
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10mg
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文献和实验mlLB,15 lbf/in2 高压灭菌20min,稍冷却,制备平皿。3. 20%麦芽糖:麦芽糖20g,加ddH2O 至100ml,0.22μm滤膜过滤。4. SM液:NaCl 5.8g,MgSO4?7H2O 2g,1M Tris?CL(PH7.5)50ml,2%明胶 5ml,加ddH2O 至1000ml。15lbf/in2高压灭菌20min。5. RNase A 10mg/ml,TE配制,沸水浴15min,分装后贮存于-20℃。6.DNase I 10mg/ml,TE配制,分装后贮存于-20℃
Fill-in Labeling of DNA Fragments
This protocol was designed to generate directionally end-labeled probes for DNaseI footprinting but it can be used for any application that requires end-labeled DNA probes. Solutions 10 mM dNTP Stocks Thaw100 mM stocks (NEB or Boehringer Mannheim
Fill-in Labeling of DNA : directional end label
m l 1 M MgCl2 10 mM DTT 100 0.5 mg/ml BSA 50 250ml 0.1 M DTT m l 10 mg/ml BSA m l Q store at -20℃ in 50 m l aliquotes dNTP Mix 21 3 Note: leave out the dNTP which will be used for labeling of the chosen restriction site (i.e. Procedure
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