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Team:Michigan/Pili June July
From 2010.igem.org
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Made a 500 mL batch of LB broth. | Made a 500 mL batch of LB broth. | ||
| - | *When pouring in distilled water, pour | + | *When pouring in distilled water, pour the powder a few mL at a time to avoid clumping. |
*It takes 20g of powder to make 1 L of broth. | *It takes 20g of powder to make 1 L of broth. | ||
Latest revision as of 01:15, 27 October 2010
| Sunday | Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | |
| Week 1 | - | 6/28/2010 | 6/29/2010 | 6/30/2010 | 7/1/2010 | - | - |
| Week 2 | - | - | - | 7/7/2010 | - | - | - |
| Week 3 | - | - | - | - | - | - | - |
| Week 4 | - | - | - | 7/21/2010 | - | - | - |
| Week 5 | - | - | 7/27/2010 | - | - | - | - |
6/28/2010
Kevin, Marc
Made a 500 mL batch of LB broth.
- When pouring in distilled water, pour the powder a few mL at a time to avoid clumping.
- It takes 20g of powder to make 1 L of broth.
Sterilized broth using the autoclave.
- The temperature setting on the autoclave is off by a little bit.
- Set dial 2 notches below 134°C.
6/29/2010
Kevin, Marc, Alena
Started growing E. coli K12 cultures:
- Poured 2 mL of LB broth into a Falcon tube
- Used strain of K12 from Dr. Lin's freezer
- Placed in incubator shaker for 24 hrs.
Added and inventoried supplies from Dr. Pinto's lab.
- Regular trash can be thrown out by going down one floor, then going outside to the trash bins.
- Chemical wastes must be cleaned by calling OSEH.
6/30/2010
Kevin, Marc
Cryopreserved stock of K12:
- 1 mL located in the iGEM box in the Lin lab.
- 1 mL located in the lab freezer.
7/1/2010
Kevin, Marc, Alena
Cryopreserved DH5α according to protocol procedure on 6/30/2010
- Put 1 stock in -20°C fridge in 1239
- Put the other stock in the -80°C fridge in the Lin lab
7/7/2010
Alena
Obtained genomic DNA of CFT073 E. coli strain from Dr. Mobley's Lab
- stored in -20°C fridge on ice
7/21/2010
Kevin, Marc, Alena
Met in Dude to determine sequence of fim operon. Arranged meeting with Dr. Mobley's group next Tuesday to learn more about hyperpiliation and the cloning process.
7/27/2010
Kevin, Marc, Alena
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.
In order to test how effectively the pili flocculate, we are planning to create an E. coli strain with fimE knocked out.
