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Hello, could you please provide clarification of the question you are asking? Are you asking how to create a restriction map, or perhaps how to go about experimentally determining the size of a given plasmid?
30 July 2012
the results i got are:
ecorI & PstI 3388 + 630
30 July 2012
hi. i had a practical to do and a set of questions to answer. i have done all of them except the last one which states to draw a restriction map.
30 July 2012
BamHI & PstI - 2818 and 954
BamHI & EcoRI - 3648 and 447
bamhi - 4074
ecorI - 4074
pstI - 4074
30 July 2012
4074 is the starting point; that's the length of the plasmid.
01 September 2012
Firstly remember the plasmid is a continuous circular piece of DNA. If we add a restriction endonuclease which has only 1 recognition sequence in the entire plasmid then it will only cleave the plasmid once. So the consequence of cleaving the circular DNA is to make it a linear piece of DNA. When we run this on DNA we get one fragment. From your data, if you look at the result we get when we cleave only with BamHI or EcoRI or pstl we get only one fragment in each case, and the fragments are all the same length. This means that all of these restriction endonucleases have one recognition sequence in this particular plasmid and so it does not matter where the enzyme cleaves because the product is always a linear 4074 bp fragment.
01 September 2012
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