MCELLBI C110L Lecture Notes - Lecture 6: Electric Field, Chain Termination, Bromophenol Blue
3 Dec 2020
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Intercalates into nucleic acid between stacked base: modify size/shape, can"t load infinite amount, evaluate quantitiy of dna then. So 2 enzymes won"t cut with same efficiency. So there will be incomplete digestion if one enzyme only cuts once. If the phosphate has been removed, phosphatase is not 100% effective though! You can always have reactions that will happen: ligation using t4 dna ligase, atp is usually in the buffer, but still add atp in the ligase reaction, transform in bacteria, and bacteria will fix itself. Transformation: can happen naturally (1% for bacteria, can happen artificially, cells are made to be competent to uptake exogenous dna, chemical transformation, electroporation. Chemical transformation: dna uptake probably through transient opening of gated membrane channels, cells treated with polyvalent cations such as cacl2, allows binding of dna to cell surface. Incubation at low temperature, followed by heat shock, heat and ionic rapid influx of extracellular medium: recovery period on rich, non-selective medium.
