How would you predict the size of fragments for two plasmids that were cut with
ID: 190010 • Letter: H
Question
How would you predict the size of fragments for two plasmids that were cut with EcoRI, HindIII, and EcoRI+HindIII before seeing the results of the gel?The two plasmids used were pFastBac-E(Z) and pFastBac-ESC with total base pairs of 4775 for each. This was done in lab and the photo was provided btu not sure how it helps.
Questions to be answered (please answer on a separate sheet and staple to the picture of the labeled agarose gel) 1. Predict the sizes of the fragments for each plasmid when cut by each enzyme alone and each of the double digests. pFastBac-E(Z) HindIII EcoRI HindIII/EcoRI pFastbac-ESC HindIII EcoRI HindIII/EcoRIExplanation / Answer
In case of pFastBac-E(Z), we will get following fragments
HindIII- Since the vector has only one site where HindIII can cleaves so we will get only one fragment that too of the linearized vector. So the size of the fragment will be 4775 bp.
EcoRI- Same will be the case with EcoRI Digested vector. We will get one fragment of size 4775 bp.
EcoRI/HindIII In the vector map site for EcoRI and HindIII are given in the MCS, that means that both the sites are maximum 100bp apart. So upon double digestion, we will get two fragments one of 4650 bp and one will be around 100 base pair.
Since nothing is given about the vector composition so I will take pFastBac-ESC has the same backbone as that of the parent vector pFastBacTM1. We will get following fragments:
HindIII- Since the vector has only one site where HindIII can cleaves so we will get only one fragment that too of the linearized vector. So the size of the fragment will be 4775 bp.
EcoRI- Same will be the case with EcoRI Digested vector. We will get one fragment of size 4775 bp.
EcoRI/HindIII In the vector map site for EcoRI and HindIII are given in the MCS, that means that both the sites are maximum 100bp apart. So upon double digestion, we will get two fragments one of 4650 bp and one will be around 100 base pair.
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