iGEM 2015 — Team SVCE_Chennai

# Team:SVCE Chennai/Notebook

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## Notebook

**APRIL 2015**

* **Recruitment of team members**
* **Setting up of lab with necessary equipments and chemicals**

* **Brainstorming**

**MAY- MID JUNE 2015**

* **Brainstorming**
* **University exams hence no lab work**

**MID JUNE- JULY 2015**

* **lab practice sessions**
* **confirmation of project title**

**AUGUST 2015**

**Week 1**

* **Submitted safety forms and also regarding our project forms.**

* **Preparation of LB broth and LB Agar and plates were poured.**
* **Stab culture for FtsZ was obtained from iGEM and was streaked on chloramphenicol LB plates.**
* **Abstract and title of our project was uploaded.**

* **Prepared chemicals and buffers for plasmid isolation.**
* **FtsZ was innoculated in LB broth.**
* **Pure culture of Pseudomonas aeruginosa and BL21 strain was obtained.**

* **Sequences for gBlocks were constructed for ordering.**

**Week 2:**

* **Plasmid isolation was performed for protocol optimisation.**

* **Result was negative and so the isolation was performed again.**
* **Elution of DNA was done but results were not satisfactory.**
* **Media (SOB and SOC) were prepared for competent cell preparation.**
* **Positive results were obtained for elution of DNA and protocol was optimised.**
* **Innoculated BL 21 in SOB.**
* **Presentation and questionnaire based on our project was prepared to educate the students in different colleges.**
* **Logo designing was initiated.**

**Week 3:**

* **Growth curve for Pseudomonas aeruginosa was done and results were analysed.**
* **Growth curve for BL 21 was done.**
* **Plasmid isolation of pET24a was performed.**
* **Competent cell of BL 21 was made and protocol was optimised.**
* **Seed stock for BL 21 competent cell was made.**
* **Transformation was performed.**
* **Primers required for our project were designed and ordered.**
* **For Human Practise questionnaire for hospitals were prepared.**
* **Presentation was done in Arulmigu Meenakshi Amman College and in Prathyusha Institute of Technology and Management.**
* **Dr. Kannan from Saveetha Medical College was interviewed regarding antibiotic resistance.**
* **Logo design was reviewed and changes were made.**
* **Approached various funding agencies.**
* **Collaboration with Uppsala was confirmed.**

**Week 4:**

* **BL21 was subcultured.**
* **Transformation efficiency of pET 24a in E coli BL21 competent cells was calculated.**
* **Chloramphenicol plates were prepared for pSB1C3 amplification.**
* **Methicillin Resistant Staphylococcus aureus were obtained.**
* **Final safety forms were filled.**
* **First step was taken for wiki page and its theme was discussed.**
* **Responses for various questionnaire was collected and a statistical analysis was carried out.**
* **Results were presented to the faculty members of our department.**
* **Three doctors from reputed medical institutions were interviewed regarding the development of antibiotic resistance and formation of biofilms in hospital environments.**

**SEPTEMBER 2015**

**Week 1:**

* **gBlocks of ftsZ was obtained.**
* **PCR amplification of the ftsZ was done.**
* **Results were found to be negative.**
* **Hence they were troubleshooted.**
* **Biochemical tests were performed for Staphylococcus and Pseudomonas. A presentation was done during the national level symposium called OMICS and created an awareness among various other college students.**

**Week 2:**

* **gBlocks of thuricin was obtained.**
* **PCR amplification of the gene was done.**
* **This amplified gene was cloned into pSB1C3.**
* **Growth curve for Staphylococcus aureus was done.**
* **Human practices team presented in Madha Engineering College and SBOA.**

**Week 3:**

* **Got bactofencin gBlocks and PCR amplification was done.**
* **Then cloning was done in pSB1C3.**
* **Cloned Thuricin and Bactofencin were submitted as parts.**
* **Public interview regarding antibiotic resistance was taken.**
* **Descriptions were uploaded in wiki page.**

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Sri Venkateswara College of Engineering, Chennai