Team:UGA-Georgia/Notebook

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Notebook

May

Week 1

Inoculation and frozen stock of colonies, Mutant frozen stocks in freezer were separated into two boxes tested and untested. Determined colonies for sequencing and DNA purification was conducted on these colonies.

Week 2

Formate media was prepared, Oxygen Exposure protocol Parts 1 and 2 were conducted on colonies inoculated in the previous week, Fluorescence measurements were taken of the colonies.

Week 3

PCR and Gel Electrophoresis was conducted on the colonies that were inoculated during previous week plan to troubleshoot variation was developed. Inoculation was conducted on more mutants and frozen stock was also made

Week 4

The mutants that were inoculated during the previous week had oxygen exposure protocol conducted on them however they were resuspended improperly, so data could not be used.

June

Week 1

- A gel was run on protein products from previous week experiment to determine what colonies should be inoculated again. Transformation of M3’s and M5’s occurred. 10 colonies were sequenced. Inoculation procedure for colonies certified from previous weeks gel and for other uncertified colonies occurred.

Week 2

Cell pellets were frozen for colonies that grew quickly. Oxygen exposure protocol was conducted on all colonies that had OD above .7. Media was made. Gel Electrophoresis and PCR was conducted on colonies that had protein extracted in previous week.

Week 3

Determined colonies for sequencing and DNA purification was conducted on these colonies. Inoculation of multiple colonies also occurred and two bench stocks of S0001 were subculture by transformation team. Agar media was also made. Frozen stocks were made and mutants were inoculated.

Week 4

- UTR Designer data gathering session and media was made. Inoculation was conducted and DNA sequencing results were determined. S001 were inoculated for transformation.

July

Week 1

Transformation of new mutants occurred. Oxygen Exposure Part 2 on the most recent triplicates had to conduct freeze/thaw of samples in pipes to lyse cells. Sequencing results were analyzed.

Week 2

Samples from the previous week were taken to the plate and fluorescent data was gathered. Data was gathered for Metabolic Modeling. Gel electrophoresis of PCR products that were made from last week’s protein extraction also occurred.

Week 3

Inosculation of new transformation and Oxygen procedure protocol occurred gels were ran of new tranformants.

Week 4

Subcultutured colonies using previously inoculated colonies that had gone into cell death. multiple gels were ran. Shipped parts for interlab project.

August

Week 1

Researchers had a meeting to determine the goals fo the team and Wiki. Shipped parts for interlab project

Week 2

PCR and gels were ran.

Week 3

3 DNA was purified for sequencing and sequencing data was analyzed

Week 4

Gel electrophoresis occurred for the parts.