IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/08

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Gel Extraction and New Digests

  • The Mcherry plate posed a few problems with enzymes, but the cuts had been made properly. It showed that the P site does not cut properly and usually results in partial digestions.
  • Made a new game plan for getting a Kanamyacin cell with three plasmids, RBS, TES, and MCherry.
  • Did a gel extraction of MCherry with X and P sites cut and froze that. Also cut out the MCherry with E and S and X and S sites but stored those with gel, and didn't finish extraction (to be used later if necessary).
  • Attached below is the plan, and the "Tuesday" part of the plan was completed as well.
  • Tuesday: Digest TesA at X and P sites and RBS at S and P and run overnight
  • Wednesday: Gel: Run in gel and extract TesA part without backbone and RBS linear plasmid with backbone.

Ligate: TesA part into RBS backbone part 

E|X| RBS | S| X| TESA | P| P|

Transform: Transform the plasmids into cells (mix plasmids with competent cells) Plate Cells

  • Thursday




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