When joining two or more DNA fragments a researcher can perform a number of manipulations to the junction of the sequences. a). Draw the structure of each end of a DNA fragment that was produced by an EcoRI restriction enzyme digestion; include in your diagram those sequences remaining on the DNA from the EcoRI recognition sequence.
The structure of the DNA containing restrictiction endonuclease site is: 5 3 3" .5 The given DNA hasrestiction sites of EcoRI (in bold).
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b). Draw the structure of the DNA molecule resulting from the reaction of this end sequence with DNA PolymeraseI and the four deoxynucleoside triphosphates.
c). Draw the sequence produced at the junction that arises if two ends the the structure derived in (b) are ligated.
d). Draw the structure produced if the structure derived in (a) is treated with a nuclease that degrades only single-stranded DNA.
e). Draw the sequence of the junction produced if the end of (b) is ligated to an end with structure (d).
f). Draw the structure of the end of a linear piece of DNA that was produced by a PvuII restriction enzyme digestion as in (a).
g). Draw the structure of the junction produced if the end with structure (B) is ligated with the end of structure (f).
h). Suppose you can synthesise a short duplex of DNA fragment with any sequence you desire. With such a fragment and the procedures outlined in (a) – (g), design a protocol that will remove an EcoRI restriction site from a DNA molecule and incorporate a new BamHI restriction site at approximately the same location.