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Team:Cornell/Notebook/Nah operon
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June
Project Description
The purpose of this subproject is to make the nah operon biobrick compatible for the use of future iGEM teams. The nah operon is an operon that converts napthalene into salicylate. In our system, this will allow napthalene to be detected by the salicylate reporter. There are three standard cutsites internal to the operon which we are removing - the biobrick compatible piece could be used by future teams in bioremediation projects.
June 10th-16th
June 13th, Wednesday
Transformed pS
Set up PCR for four fragments of the nah operon as well as the pBMT-1 backbone, using primers designed to mutate cut-sites concurrently with Gibson Assembly. Also set up a PCR for the entire nah operon; if Gibson Assembly fails, but we are able to PCR the entire 10kb nah operon using Phusion polymerase, an alternate method of mutation using Phusion will be pursued.
June 14th, Thursday
June 17th-23rd
June 19th, Tuesday
- Ran Gibson Assembly of nah operon fragments into pBMT-1 backbone and transformed into DH5a electrocompotent E. coli cells.
- Set up PCR to amplify the Gibson Assembly products.
- Work done by: Swati and Dylan
- Tested electroporator in Weill, transforming into DH5a electrocompotent cells borrowed from the DeLisa lab.
- Word done by: Claire, Dylan, and Steven
June 20th, Wednesday
- Set up a digestion of the PCR of Gibson Assembly products and ran the digested products on a gel to see if Gibson worked.
- We'll also check to see if we got colonies from the transformation then.
- Work done by: Swati and Dylan
June 24th-30th
June 27th, Wednesday
- Ran digest of gibson-assembled nah operon on gel
====June 28th, Thursday====
