Team:Cornell/Notebook/Nah operon

From 2012.igem.org

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===[[Team:Cornell/Notebook/Nah_operon/July15|July 15th-21st]]===
===[[Team:Cornell/Notebook/Nah_operon/July15|July 15th-21st]]===
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===June 17th-23rd===
 
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====June 17th, Sunday====
 
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*Successful PCR of pBMT-1 [[Gel Purification | gel purified]].
 
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====June 19th, Tuesday====
 
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*Ran [[Gibson Assembly]] of nah operon fragments into pBMT-1 backbone and [[Transformation | 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: Dylan and Swati''
 
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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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*Three colonies on plates of DH5a transformed with Gibson Assembly product. Made [[liquid cultures]] to miniprep and sequence.
 
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**''Work done by: Dylan and Swati''
 
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====June 22nd, Friday====
 
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*[[Miniprep | Miniprepped]] directly transformed Gibson Assembly product for sequencing using the the Gibson nah1F and nah4R primers (each w/ 20 bp overhangs).
 
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*Ran undigested miniprep with [https://2011.igem.org/Team:Cornell/Protocols gel electrophoresis], looking for large bands corresponding to supercoiled DNA. The gel shows distinct bands for all three lanes. We interpreted this to mean that we got product. Submitted for [[sequencing]]
 
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::[[File:Cornell2012_0621_Gibson-entire-plasmid.jpg|600px]]
 
===June 24th-30th===
===June 24th-30th===

Revision as of 21:13, 3 July 2012

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Contents

The purpose of this subproject is to make the nah operon biobrick compatible for the use of future iGEM teams. The nah operon codes for a pathway that converts naphthalene into salicylate. In our system, this will allow naphthalene to be detected by the salicylate reporter. There are three standard cut sites internal to the operon which we are removing - PstI, and two NotI sites. This biobrick compatible piece could be useful to future teams in projects that involve bioremediation or detection of naphthalene.

June

June 10th-16th

June 17th-23rd

June 24th-30th

July

July 1st-7th

July 8th-14th

July 15th-21st

June 24th-30th

June 27th, Wednesday

Rafael and Dylan making an agarose gel.

====June 28th, Thursday====

Retrieved from "http://2012.igem.org/Team:Cornell/Notebook/Nah_operon"