Team:TzuChiU Formosa/Notebook

From 2009.igem.org

Revision as of 13:12, 21 October 2009

Notebook

Meeting

2009.03 　IGEM recuiting Poster Posted on Campus

People from different departments grouped together after they saw the poster.

2009.03.14 　Experience sharing

NYMU-Taipei 2008 IGEM team came to TCU and shared Experience with students and faculty of TCU

2009.05.07 　1st meeting

Advisors introduced iGEM

2009.05.15 　2nd meeting

Groups formed and started brain storming. 12 ideas were presented for discussion, Please see the "Brain Storm" section of our wiki for these 12 ideas.

2009.05.28　 3rd meeting

Brain storming and two projects out of 12 were voted out

2009.07~2009.09 　English courses

English conversation courses given on Wednesdays, Thursdays.

2009.08.09 　Visiting friends from Japan iGEM teams

Visiting students from University of Tokyo, and Chiba University, Japan iGEM teams.

2009.08.31 　4th meeting

Progress report of the two groups.

2009.09.14 　5th meeting

chose 1 team out of 2 .

2009.09.19 　6th meeting

Reference searching.

2009.09.23 　7th meeting

Reference searching and experimental designs.

2009.09.30 　8th meeting with advisers

Reference searching and experimental designs. Discuss with advisors

2009.10.2 　9th meeting

Duties assigned. Started wiki editing

2009.10.11 　10th meeting

Progress report and upload "project", "protocals" to wiki.

2009.10.15 　11th meeting

Progress report.

2009.10.17 　12th meeting

Upload results, discussion, notebook to wiki.

2009.10.20 　13th meeting

Progress report and modification of wiki with advisors. 　

Experiments

2009.10.06 　OmpC promoter transformation.

Plasmid pSB1A3 biopart from 96-well plated was dissolved in 10 ul PCR water, 2 ul was used for plasmid transformation into DH5α competent cells.

2009.10.08　Positive clone selection and plasmid isolation.

4:30 pm ~ 4:50 pm Pick single bacterial colony that had been transform with OmpC promoter containing plasmid and inoculate with 3ml LB medium containing Ampicillin and grow overnight (16-20 hours) in a 37℃ shaker incubator.

2009.10.09　Colony PCR to confirm OmpC promoter and plasmid isolation.

2:00 pm ~ 5:00 pm 10 ul overnight culture was used for colony PCR. Briefly 10 ul overnight culture was centrifuged at 13,000 rpm for 10 min, supernatant was removed, 500 ul autoclaved water was added fllowed by 100C incubation for 20 min and chilled on ice. 5 ul was used for colony PCR in a total 20 ul reaction with OmpC promoter specific primers. 5 ul of the PCR products was used for agarose gel electrophoresis to verify the positive clone.

2009.10.11　Prepare LB plates

8:30 am ~12:00 noon 500 ml LB-agar was prepared and autoclaved. When LB-agar was ready (temperature cold enough to hold, about 60°C), 500 ul ampicillin (50 mg/ml) and 500 ul kanamycin (50 mg/ml) were added and 14 LB-agar plates were prepared.

4:10 pm ~ 4:30 pm pSB1A3 preparation Three single colonies containing pSB1A3 plasmid were inoculated in 5ml LB medium containing 7μl ampicillin (50 mg/ml) and grow overnight (16-20 hours) in a 37℃ shaker incubator.

2009.10.11　Stock bacterial preparation and plating new plates for future used.

4:30 pm ~ 4:50 pm Two glycerol stock were prepared as follow : 0.5 ml overnight culture was added into 0.5 ml 20% glycerol stock in a 2 ml eppendorf tube, mix well and kept at -80C.

4:35pm~4:50pm Select two OmpC promoter transformed single colonies and inoculate 5ml LB contain 7μl Ampicillin and grow overnight (16-20 hours) in a 37℃ shaker incubator.

2009.10.12 　T-A cloning.

2009.10.13　T-Atransformation.

2009.10.14　Cloning PCR.

2009.10.16 　T-A cloning.

2009.10.20 　Digestion, Gel extration

2009.10.21 　Gel extration

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