Lecture 5 - Lecture5

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Lecture 5 Nucleic acid hybridization - artificially denaturing double stranded DNA to allow the single stranded DNA to anneal with another sequence significance of doing this: In order to know what region the DNA is from, the amount of RNA produce, etc 1 strand is called the probe because it has a quantifiable tag the tag exists so that the researcher can visualize where the desired comple- mentary sequence can be found in the genomic sequence in the past the probe was made by using radioisotope, presently due to restric- tions, the present methods use antibody or fluorescent The T m is the temperature at which half the double stranded DNA has denatured stability of hybrid - T m bases can’t hybridize, the H-bond flips out and H-bonding occurs with the water. This occurs if the T m is not known for the DNA being used. At the temperature is raised, T m = half single and half double stranded DNA The more GC content in the DNA the greater the stability and the greater heat re- quired to denature DNA Have to vary parameters for T m so compatible for appropriate hybridization can oc- cur Need to find the ideal temperature so that the hybrid can form and not stay in a denatured state Temperature has to be low enough for the hybrid to occur human genome denatures at 89 o C Genetic code is degenerate (multiple codons exist for the same amino acids) Cell constantly denaturing and annealing DNA, The cell must have an appropriate amount of GC vs AT content so that the DNA is not denatured easily as well as it has to be such that the minimum amount of GC
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Lecture 5 - Lecture5

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