Team:SASTRA Thanjavur/Notebook
Notebook
Throughout our struggles and hard work, people came and people left, but one thing stood by our side - Our notebook. Errors, mistakes and perfect results - Our notebook has seen it all, documenting our entire journey, every step of the way. Here it is - Hope you enjoy reading it as much as we enjoyed writing it.
| Date | Description |
|---|---|
| 02-July | Preservation of E.coli MG1655 Culture for plasmid transfection |
| 09-July | Plasmid isolation by alkaline lysis method |
| 13-August | Planning and protocol discussion for PCR experiments |
| 15-August | Analysis of DNA Distribution kit for parts characterization - to study the effect of pH on GFP expression on the selected plasmid- pSB1C3 |
| 22-August | High efficiency transformation of pSB1C3 using NEB C2987I - protocol design |
| Preparation of experimental requirements and its sterilization | |
| Requirements - Distilled water for preparation of antibiotics, LB agar, eppendorf vials & tip boxes | |
| 23-August | Implementation of DNA resuspension protocol |
| 24-August | Plasmid transformation into the competent cells provided |
| 26-August | Repetition of transformation experiment due to poor transformation efficiency. |
| 1st PCR experiment - Toehold for miR20a | |
| 28-August | Preparation of experimental requirements and its sterilization |
| Requirements - Distilled water for preparation of antibiotics, LB broth, LB agar, eppendorf vials & tip boxes | |
| 29-August | Subculturing of the formed colonies of plasmid transformed competent cells |
| 30-August | Repetition of transformation experiment - We noticed some precipitation in the SOC broth and decided to repeat the experiment using LB broth and decreasing the RPM to 150. |
| 07-September | Fluorescence analysis of DH5 Alpha and the transfected cells (Strain 8) after 48 hours of incubation |
| 09-September | Trial 2 - Inoculation of colonies from Strain 8 and DH5 Alpha into freshly prepared LB broth |
| 10-September | Fluorescence analysis of the samples inoculated on 09-Sept, after 24 hours of incubation |
| 11-September | Trial 3 - Inoculation of colonies from Strain 8 and DH5 Alpha into freshly prepared LB broth |
| Fluorescence analysis of the samples inoculated on 09-Sept, after 6 hours of incubation | |
| 12-September | Performed OD normalization to set the number of cells for fluorescence measurement after 24 hours |
| Fluorescence analysis of the samples inoculated on 09-Sept, after 24 hours of incubation | |
| 2nd PCR experiment - Toehold for miR 21, miR 29 | |
| Inoculation of Strain 8 in different CAMP (chloramphenicol) concentrations | |
| 14-September | Performed OD normalization at 595nm to normalize the number of cells for fluorescence measurement after 48 hours |
| Fluorescence analysis of the samples inoculated on 12-Sept, after 48 hours of incubation | |
| Planning and protocol discussion for pH study | |
| 15-September | Preparation of LB broth media in varying pH (5, 6, 7, 8, 9) and sterilized |
| Preparation of 20 µg/mL of CAMP stock | |
| 16-September | Trial 1 - Inoculated Dh5 Alpha and Strain 8 in varying pH media (with CAMP for Strain 8) |
| Preperation of LB broth media in varying pH (5, 6, 7, 8, 9) and sterilized | |
| 17-September | Trial 1 - Fluorescence study after 24 hours of inoculation |
| Trial 2 - Inoculated Dh5 Alpha and Strain 8 in varying pH media (with CAMP for Strain 8) | |
| 3rd PCR experiment - Toehold for miR20a and miR 21 (sample 1 in 3 dilution) | |
| 19-September | Trial 2 - Performed OD normalization at 595nm to set the number of cells for fluorescence measurement after 48 hours |
| Trial 2 - Fluorescence analysis of the samples inoculated on 17-Sept, after 48 hours of incubation | |
| 21-September | Preparation of LB broth media in varying pH (5, 6, 7, 8, 9) and sterlized |
| 22-September | Trial 3 - Inoculation of colonies from Strain 8 and DH5 Alpha into freshly prepared LB broth |
| 23-September | Trial 3 - Performed OD normalization to set the number of cells for fluorescence measurement after 24 hours |
| Trial 3 - Fluorescence analysis of the samples inoculated on 22-Sept, after 24 hours of incubation with LB broth as reference | |
| 24-September | Trial 3 - Performed OD normalization at 595nm to set the number of cells for fluorescence measurement after 48 hours |
| Trial 3 - Fluorescence analysis of the samples inoculated on 22-Sept, after 48 hours of incubation with LB broth as reference | |
| 4th PCR experiment - Toehold for miR20a and miR21 (1 uL sample directly without dilution) | |
| 26-September | 5th PCR experiment - Toehold for miR20a and miR21 (1 uL sample directly without dilution) |
| 03-October | Trial 1 - Fluorescein Standard graph plot for instrument calibration & GFP reference (Auto gain and gain set to 70) |
| 04-October | Trial 2 - Fluorescein Standard graph plot for instrument calibration & GFP reference (Gain set to 65, 70, 90, 100) |
| 09-October | Purification of PCR product |
| Characterization of toehold for miR-200a ( Trial run 1) | |
| 10-October | Quantification of purified PCR product |
| Characterization of toehold for miR-21 ( Trial run 2) | |
| 14-October | Characterization of toehold for miR-21 ( Concentration and specificity Study). Performed by the sensor team. |
| 16-October | Characterization of toehold for miR-20a ( Concentration and specificity Study) Performed by the sensor team. |
The "blueprint" of the House of Wetlab and Sensors can be accessed here for your reference.