ApE, a Plasmid editor

Examples of how to use ApE:

 

Start ApE

 

Paste the sequence of Lambda DNA (from SCLResources>sequences)
(note:
in the example pictures shown, the lambda sequence files was not saved, so Lambda DNA is called New DNA..)

note:
in the bar below the line of icons is shown the following:
position of the mouse pointer, start of a selection, length of a selection, end of a selection.

 

 

Go to the Enzyme Selector:

 

and click on BamHI and EcoRI; both enzymes show to have (5) recognition sites in Lambda DNA.

 

Click the Digest button to get the digestion result

 

Hover the mouse arrow over the bands, map or text, to get additional info:
e.g.: the 3531 bp fragment runs from the EcoR I site at position 44972 to the end (cos-end) of Lambda DNA

and the other end fragment is 5504 bp long and runs from the start (cos-end) to the BamH I site at position 5505.

 

When you click on a band (or fragment) in the digest window, the corresponding part of the sequence in the main ApE window is highlighted!
When you e.g. click on the 1120 fragment,

the sequence of this fragment is highlighted, with the EcoR I (gaattc) site at one end (position 21226) and the BamH I site (ggatcc) at the other end (position 22346)

 

 

In solution, some of the lambda DNA molecules are joined at their cos-ends, to form the cos-site. Those circular (or multimeric) Lambda DNA molecules do not give the end fragments of 5504 and 3531 basepairs, but a combination of those two fragments, giving one big fragment of 9035 basepairs.
To show this using ApE, change the linear/circular button to circular,

and digest again using BamH I and EcoR I.
Hovering over the end fragments show this time one fragment of 9035 bp.

 

 

To show the two Hind III sites in the insert of pGTl3758, highlight the 3758 fragment in the digest window.
It shows that the 3758 bp fragment runs from BamH I (at position 22346) to EcoR I (at 26104):

Clicking on the 3758 fragment highlights it's sequence in the main window, with EcoR I (gaattc) at one end

and BamH I (ggatcc) at the other end.

 

To find the two Hind III sites, used in the pGTl3758DH construction, select HindIII (de-select any other enzyme..) in the enzyme list, with Selection checked (see mouse arrow in the picture below)

and click the Highlight button. The two Hind III sites within the are shown in red.

Clicking the Digest button in the Enzyme Selection window shows the positions of the Hind III sites (23130 and 25157) and the length of the Hind III fragment (i.e. the deletion in pGTl3758DH !)