Team:Cornell/Notebook/Nah operon

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==June==
 
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===Project Description===
 
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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.
 
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===June 10th-16th===
 
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====June 13th, Wednesday====
 
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Transformed pS
 
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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.
 
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====June 14th, Thursday====
 
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===June 17th-23rd===
 
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====June 18th, Monday====
 
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Tomorrow, Claire et al. will also be testing out the electroporator in Weill so that we can transition to the BME lab space. This will be happening after the Gibson stuff in finished in Olin. So... you can venture to Weill if you are excited about electroporation.
 
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====June 19th, Tuesday====
 
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Ran Gibson Assembly of nah operon fragments into pBMT-1 backbone and transformed into DH5a electrocompotent ''E. coli'' cells.
 
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Set up PCR to amplify the Gibson Assembly products.
 
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''Work done by: Swati and Dylan''
 
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====June 20th, Wednesday====
 
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Set up a digestion of the PCR of Gibson Assembly products and ran the digested products on a gel to see if Gibson worked.
 
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We'll also check to see if we got colonies from the transformation then.
 
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''Work done by: Swati and Dylan''
 
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===June 24th-30th===
 
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====June 27th, Wednesday====
 
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*Ran digest of gibson-assembled nah operon on gel
 
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====June 28th, Thursday====
 

Latest revision as of 06:40, 26 October 2012