This page contains a weekly account of the experiments and activities conducted in the laboratory.
Week 1
Specific buffers were made and autoclaved. Prepared antibiotic solutions and poured plates. DH5-α competent cells were prepared, and PCR mixes were made for 4 different DNA sequences and 5 different controls. Transformant bacteria (RFP) were created. E. coli was streaked.
Week 2
LB was autoclaved, and plates were spread. Absorbance values for E. coli and B. cereus was measured. Antibiotic solutions of Ampicillin and Chloramphenicol were made.
Week 3
Gel Electrophoresis for CcaS-PCR, was performed. PCR was performed for the miniprepped products of ECFP-1 and ECFP-2. Bba_K3165039 was ordered. Bba_K3165041 and Bba_K3165036 was ordered for CcaS system. For Opto-T7 system, Bba_K3165050, Bba_K3165052 and Bba_K3165054 was ordered.
Week 4
Gradient PC/h6>Rs for PMag-gP2 was carried out with different initial reagent concentrations. PCR for CcaS system was also performed and PCR, for CcaR and Cyan was performed. PCRs for Opto-T7 NMag and touchdown PCR for CCaR and Cyan were performed. Miniprep for pBS1C was performed.
Week 5
Restriction digestion of pBS1C and pBS4S was performed. Cloning trial for Opto-T7 was performed. Restriction digestion was performed on the following plasmids: pBS4S, pBS1C and PMag. SDS-Page for ECF-P was performed. Double digest with EcoRI and PstI was performed. Midiprep for pSB1C3 was performed. pBS4S and pBS1c were ligated.
Week 6
Digestion on last week's midiprep was performed. First attempt for ligation of pSB1C3 with CcaR was performed. pBS1C and pBS4S were restriction digested for E and P.
Week 7
Ligation was attempted for the second time. Miniprep was performed on 8 of the chosen colonies which was obtained for the aforementioned ligation. Gel extraction for pBS1C and pBS4S was performed. Double digestion for CcaR ligation for 18 colonies was carried out.
Week 8
Gel electrophoresis was carried out, from 10 of the CcaR-digested colonies and PCR was carried out on the DNA from the colonies. ECFP characterization was attempted.
Week 9
PCR for mCherry and i2mCherry(Leu) was carried out. The DNA concentration for the above parts were found out through NanoDropTM. A gradient PCR for improvement of parts was done.
Week 10
The digested pSB1C3 from RFP-Biobrick and the RFP fragment were purified by the standard agarose gel-extraction procedure. Ligation for improvement-part was carried out.
Week 11
The ligated product was transformed. Digesition for pSB1C3(ECFP), miniprepped, i2mCherry(Ile) PCRed product and i2mCherry(Leu)-pSB1C3 ligated product was obtained.
Week 12
Improvement of pSB1C3 for restriction sites E and S was carried out. Gel extraction was carried out in an attempt to extract the required plasmid.
To view a detailed account of our laboratory activities, download the PDF attached below.
