This web page was produced as an assignment for an undergraduate course at Davidson College.

On new page: In order to determine if the plasmid is carrying the insert in the correct orientation, it is necessary to cut the plasmid with a specific restriction enzyme that will run different lengths when electrophoresed depending on the orientation of the insert. In clearer terms, if the insert is in the correct orientation, it will give one set of specified lengths, and if it is in the reverse orientation, the gel will run a set of different specified lengths.  In my experiment (at least in the one on this web page) the reverse orientation is determined to be present if the forward orientation lengths are not present on the gel. I understand this is a negative control (gasp!) but I was unsure of how to calculate the specified lengths in the reverse direction.

Model:  ID[X] - Restriction enzyme used for digestion
(If the insert is in the forward direction the lengths on the gel will be) ~x bp and ~y bp
 
IDH1 - Kpn I
~340 bp and ~4265 bp
 
IDH2 - Eco RV
~802 bp and ~3808 bp
 
IDP1 - Bgl II
~90 bp and ~4695 bp
 
IDP2 - Bgl II
~508 bp and ~4231 bp
 
IDP3 - Cla I
~508 bp and ~4243 bp

 

This page is maintained by Nicole Hesson.

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