Team:BU Wellesley Software/Notebook/TraciNotebook

From 2011.igem.org

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Revision as of 20:04, 15 July 2011

Mentoring Notes

The protocols used by this team can be found at:

Protocols

Finally iGEM kick off week arrived! I finished the BioBasics lectures with the entire team (both BU and Wellesley members were present). Following Orit's suggestion, I had the students break into teams and explain figures from a paper to the entire team to ensure students really did understand the concepts and results discussed in the paper.

Training for the wet team continued on Tuesday and Wednesday. During lab meeting on Wednesday, tasks were assigned to groups for building the first constructs the team will need to make (promoter-rbs-xFP-terminator).

These tasks included the following:

1. Choosing promoters, ribosomal binding sites and fluorescent proteins to build 4-part transcriptional units to generate fluorescence

2. Transforming the chosen BioBrick parts into Top10 competent cells

3. Miniprepping and carrying out restriction digests on these parts

4. Gel electrophoresis and extraction to confirm restriction digests

5. Ligations with the cut backbones and inserts

6. Repeat 1-5 until transcriptional units are made

Week 2: 6/13-6/17/2011

I was at SB5.0 from 6/14-6/18 and was mentoring remotely via email.

Week 3: 6/20-6/24/2011

I was away 6/22-6/25 and mentored remotely via email.

Week 4: 6/27-7/1/2011

Kyle, Alberto and I met with Jenhan and Rishi to discuss the Wet Lab needs for Batterboard. We discussed creating a different "template" for saving tubes instead of saving data in 96-well plates. We recommended creating an 81-grid box or rack in lieu of the 96-well plate since our current work has been done in 1.5mL tubes and not plates. With this 81-grid rack in mind, we want three "types" of these racks to choose from - a Plasmid rack, a Gel Extraction rack and a Frozen Stock rack to store our various types of samples.

I also worked with Swapnil one afternoon to generate and test a new liquid handling class for the robot so we can accurately pipet enzymes in glycerol.

Week 5: 7/4-7/8/2011

Given the lack of success of our transformations with ligations, I made a new batch of competent cells following the Anderson protocol using E. coli TOP10 cells. Over 200 tubes of 200uL aliquots of cells were prepared and frozen at -80C.

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+Given the lack of success of our transformations with ligations, I made a new batch of competent cells following the Anderson protocol using ''E. coli'' TOP10 cells. Over 200 tubes of 200uL aliquots of cells were prepared and frozen at -80C.
-I made new competent cells following the Anderson protocol.
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