When double-stranded DNA is heated, the two strands separate into single strands in a process called melting or denaturation.The temperature at which half of the duplex DNA molecules are intact and half have melted is defined as the Tm.
A.Do you think Tm is a constant, or can it depend on other small molecules in the solution? Do you think high salt concentrations increase, decrease, or have no effect on Tm?
B.Under standard conditions, the expected melting temperature in degrees Celsius can be calculated from the equation Tm = 59.9 + [0.41 × %(G + C)] − (675/length of duplex).Does the Tm increase or decrease if there are more G + C (and thus fewer A + T) base pairs? Does the Tm increase or decrease as the length of DNA increases? Why?
C.Calculate the predicted Tm for a stretch of double helix that is 100 nucleotides long and contains 50% G + C content.
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