Difference between revisions of "Team:UNOTT/Design2"

Line 90: Line 90:
 
<p style="text-align: center;"><span style="color: #ffffff;">___________________________</span></p>
 
<p style="text-align: center;"><span style="color: #ffffff;">___________________________</span></p>
 
<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/2017/5/56/UNOTT2017-plasmiddescript.png" alt="" width="800" height="400" />&nbsp;</p>
 
<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/2017/5/56/UNOTT2017-plasmiddescript.png" alt="" width="800" height="400" />&nbsp;</p>
 +
 +
These methods are used to create two plasmids, both of which to some extent are randomly assorted. An sgRNA plasmid which complements the dcas9, and a reporter plasmid which expresses the randomly inhibited and produced reporter FP's along with a dcas9.
  
  

Revision as of 00:16, 14 September 2017

.

KEY PLASMID DESIGN

 

 

Random Brick formation (Components of plasmid)

___________________________

 

 

 

These bricks are formed from a random soup of characterised promoters "P", a reporter gene fluorescent protein, and a random terminator "T". This uses BSA I sites already digested previously into the DNA in order to ligate randomly in the correct order to create a random, yet purposeful and functional fluorescent signal.

Brick stitching

___________________________

 

These bricks are then stitched together via amplifying each randomly assembled brick through common amplification sites and then cutting them using a set of restriction enzymes which give each plasmid a specific order of bricks, depending on which are cut and then ligated together. As shown.

Plasmid Design

___________________________

 

These methods are used to create two plasmids, both of which to some extent are randomly assorted. An sgRNA plasmid which complements the dcas9, and a reporter plasmid which expresses the randomly inhibited and produced reporter FP's along with a dcas9.