You add 3.0 µg of plasmid DNA to a QIAprep spin miniprep purification column. You perform the binding in a binding buffer at pH 6.8. What portion (%) of the 3.0 µg of plasmid DNA would you expect to bind to the column in these conditions? What portion (%) of the bound plasmid DNA would you expect to elute using elution buffer at pH 8.5? Make the following assumptions: • Solid phase silanol groups can have a very wide range of pKas. For the purpose of this problem, assume the silica gel functional group (silanol) in the DNA purification column matrix has a pKa of 7.5. • If a DNA molecule and column matrix functional groups have the same charge state, then that DNA molecule will be repelled and will not bind to the column. If they do not have the same charge, binding occurs.
You add 3.0 µg of plasmid DNA to a QIAprep spin miniprep purification column. You perform the binding in a binding buffer at pH 6.8. What portion (%) of the 3.0 µg of plasmid DNA would you expect to bind to the column in these conditions? What portion (%) of the bound plasmid DNA would you expect to elute using elution buffer at pH 8.5?
Make the following assumptions:
• Solid phase silanol groups can have a very wide range of pKas. For the purpose of this problem, assume the silica gel functional group (silanol) in the DNA purification column matrix has a pKa of 7.5.
• If a DNA molecule and column matrix
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