Team:Michigan/Pili Expression

From 2010.igem.org

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!Week 1
 
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|[[Team:Michigan/Pili_Expression#6/28/2010|6/28/2010]]
 
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|[[Team:Michigan/Pili_Expression#6/29/2010|6/29/2010]]
 
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|[[Team:Michigan/Pili_Expression#6/30/2010|6/30/2010]]
 
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=Pili Expression Team=       
=Pili Expression Team=       
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This team includes [[User:MarcSinger|Marc Singer]], [[User:kevijose|Kevin Joseph]], and [[User:shanwu|Alena Wu]].
This team includes [[User:MarcSinger|Marc Singer]], [[User:kevijose|Kevin Joseph]], and [[User:shanwu|Alena Wu]].
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==6/28/2010==  
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==Monthly Notebooks==
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Made a 500 mL batch of LB broth
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In order to conserve page space, we have split our notebook according to months.
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*When pouring in distilled water, pour a few mL at a time to avoid clumping.
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*It takes 20g of powder to make 1 L of broth
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Sterilized broth using the autoclave
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[[Team:Michigan/Pili_June_July|June/July]]
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*The temperature setting on the autoclave is off by a little bit
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*Set dial 2 notches below 134°C.
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==6/29/2010==
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[[Team:Michigan/Pili_August_September|August/September]]
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Started growing E. coli K12 cultures
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*Poured 2 mL of LB broth into a Falcon tube
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*Used strain of K12 from Dr. Lin's freezer
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*Placed in incubator shaker for 24 hrs.
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Added and inventoried supplies from Dr. Pinto's lab.
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[[Team:Michigan/Pili_October|October]]
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*Regular trash can be thrown out by going down one floor, then going outside to the trash bins.
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*Chemical wastes must be cleaned by calling OSEH.
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==6/30/2010==
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Cryopreserved stock of K12
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*1 mL located in the iGEM box in the Lin lab.
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*1 mL located in the lab freezer.
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==7/1/2010==
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Cryopreserved DH5α according to protocol procedure on 6/30/2010
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*Put 1 stock in -20°C fridge in 1239
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*Put the other stock in the -80°C fridge in the Lin lab
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==7/7/2010==
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Obtain genomic DNA of CFT073 E. coli strain from Dr. Mobley's Lab
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*stored in -20°C fridge on ice
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==7/21/2010==
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''Kevin, Marc, Alena''
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Met in Dude to determine sequence of ''fim'' operon.
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Arranged meeting with Dr. Mobley's group next Tuesday to learn more about hyperpiliation and the cloning process.
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==7/27/2010==
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''Kevin, Marc, Alena''
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Met with Chris Alteri from Dr. Harry Mobley's research group to discuss the best route to hyperproduce the pili. Chris recommended that we create a plasmid by cloning FimB into pBAD, and then inserting that plasmid in MG1655. In theory, that should activate flocculation in the E. coli, inducible by arabinose. Chris was able to give us the procedures for creating a plasmid with FimB, as well as the procedures for knocking out a gene.
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In order to test how effectively the pili flocculate, we are planning to create an E. coli strain with fimE knocked out.
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==8/7/2010==
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''Kevin, Marc, Alena''
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PCR #1
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Used a gradient from 40C to 60C for the first 3 cycles to find the optimum anneling temperature.
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All of the annealing temperatures gave a good result according to the gel.
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[[media:Lab10.pdf|8/7/10 notes]]
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==8/9/2010==
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''Kevin, Marc, Alena''
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Used a 57C degree annealing temperature to get enough DNA for the digest and ligation.
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4 out of the 5 PCR reactions worked well according to the gel.
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[[media:Lab11.pdf|8/9/10 notes]]
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==8/11/2010==
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''Kevin, Marc''
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Met with Chris, received advice for updating digest and ligation protocols.
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|width="250px" style="background-color:rgb(190,210,250);"|
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=='''In the Lab'''==
=='''In the Lab'''==
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<html>
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[[Image:pili01.png|middle|250px]]
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<iframe src="http://www-personal.umich.edu/~shanwu/wiki_photos/src/themes/classic/classic-demo_pili.html" width="100%" height="650px" scrolling="no"></iframe>
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</html>
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[[Image:pili02.png|middle|250px]]
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[[Image:pili03.png|middle|250px]]
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[[Image:pili04.png|middle|250px]]
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[[Image:pili05.png|middle|250px]]
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[[Image:pili06.png|middle|250px]]
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Latest revision as of 07:56, 26 October 2010


Michigan Header




Pili Expression Team

This team includes Marc Singer, Kevin Joseph, and Alena Wu.

Monthly Notebooks

In order to conserve page space, we have split our notebook according to months.

June/July

August/September

October

In the Lab

Retrieved from "http://2010.igem.org/Team:Michigan/Pili_Expression"