Notebook
Table of Content
| Date | Entry Title | Group | Notebook | Page |
|---|---|---|---|---|
| Aug 26, 2025 | PCR of BLIP-I | A | 1 | 1 |
| Dilution of 10× TAE Buffer to 1× and Autoclaving | A | 1 | 2 | |
| Aug 29, 2025 | Restriction Digestion of pSB1C3 | A | 1 | 2 |
| Preparation of LB agar and LB broth | B | 1 | 3 | |
| Rehydration of E.coli S17-1 | B | 1 | 3 | |
| Aug 30, 2025 | Resuspension of BLIP-II, PCR of BLIP-II, and Restriction Digestion of BLIP-I & pSB1C3 | A | 1 | 4 |
| Preparation of S17-1 glycerol stock | B | 1 | 4 | |
| Sep 2, 2025 | Gel electrophoresis of products from 8/30 (BLIP-II PCR, BLIP-I digest, pSB1C3 digest) | A | 2 | 1 |
| Subculture of E.coli | B | 2 | 2 | |
| Sep 4, 2025 | Reviving E. coli DH5α from Glycerol Stock – Plate (Part 1) | B | 2 | 2 |
| Preparation of S17-1 Glycerol Stock (from Plate) - Failed Attempt | B | 2 | 3 | |
| Sep 5, 2025 | Gel electrophoresis of BLIP-I RD, pSB1C3 RD, and BLIP-II PCR | A | 2 | 4 |
| Reviving E. coli DH5α from Glycerol Stock – Broth (part 2) | B | 2 | 5 | |
| Inoculation of S17-1 into LB Broth | B | 2 | 5 | |
| Sep 6, 2025 | Gel extraction of products made on 9/5, PCR and restriction digestion of BLIP-I and BLIP-II, gel run of both | A | 2 | 6 |
| Preparation of S17-1 Glycerol Stock (from Broth) | B | 2 | 7 | |
| Preparation of Antibiotic Plates (Amp + Chl) | B | 2 | 7 | |
| Sep 8, 2025 | Ligation and gel run of BLIP-I (PCR product made on 9/6) | A | 3 | 1 |
| Resuspension of BBa_J435300 (Distribution Kit DNA) | B | 3 | 1 | |
| Sep 10, 2025 | Gel Extraction of BLIP-I (PCR product from 9/6) | A | 3 | 2 |
| Heat-Shock Transformation of BBa_J435300 into DH5α | B | 3 | 2 | |
| Sep 12, 2025 | Ligation of BLIP-I and BLIP-II (T4 DNA ligase) | A | 3 | 3 |
| Overnight Culture of S17-1 Subculture | B | 3 | 3 | |
| Sep 13, 2025 | Preparation of Competent Cells (S17-1, CaCl2 method) | A&B | 3 | 4 |
| Sep 15, 2025 | Subculture of E. coli S17-1 (from 08/29 rehydration plate) | B | 4 | 1 |
| Sep 16, 2025 | Preparation of Competent Cells (CaCl₂ method, Attempt 1) | A | 4 | 2 |
| Preparation of LB Agar and LB Broth (no antibiotics & Amp) | B | 4 | 2 | |
| Sep 17, 2025 | PCR of BLIP-I | A | 4 | 3 |
| Preparation of Competent Cells (CaCl₂ method, Retry) | B | 4 | 3 | |
| Sep 18, 2025 | PCR of BLIP-II | A | 4 | 4 |
| Resuspension of pSB1C3-mCherry (Distribution Kit DNA - O11) | B | 4 | 4 | |
| Heatshock transformation of (DH5α + mCherry, pSB1C3) | B | 4 | 4 | |
| Sep 19, 2025 | BLIP-I gel run | A | 4 | 5 |
| PCR of BLIP-II | A | 4 | 6 | |
| BLIP-I purification | A | 4 | 6 | |
| Heat Shock Transformation (DH5α + BBa_J435300) | B | 4 | 6 | |
| Plating of Pinky (DH5α + mCherry, pSB1C3) | B | 4 | 6 | |
| Sep 20, 2025 | Gel Electrophoresis of BLIP-I & BLIP-II | A | 4 | 7 |
| Plating | B | 4 | 8 | |
| Miniprep of Pinky Plasmid (pSB1C3 backbone) | B | 4 | 8 | |
| Preparation of Selective Agar Plate | B | 4 | 9 | |
| Sep 22, 2025 | Restriction Digestion of BLIP-I, BLIP-II, and pSB1C3 | A | 5 | 1 |
| Sep 23, 2025 | Gel Electrophoresis of 9/22 Restriction Digestion Products (BLIP-I, BLIP-II, pSB1C3) | A | 5 | 2–3 |
| Resuspension of BBa_J435500 (Distribution Kit DNA - O6) | B | 5 | 3 | |
| Heat Shock Transformation (BBa_J435500 + DH5α) | B | 5 | 4 | |
| Sep 24, 2025 | Restriction Digestion and Gel Electrophoresis of BLIP-I, BLIP-II, and pSB1C3 | A | 5 | 5 |
| Subculture and Liquid Culture of DH5α + BBa_J435500 (no antibiotic) | B | 5 | 6 | |
| Sep 25, 2025 | PCR of BLIP-I | A | 5 | 7 |
| Sep 26, 2025 | Gel run of samples from 9/25 and gel extraction following the new method | A | 5 | 8 |
| Overnight Culture & Glycerol Stock Preparation | B | 5 | 9 | |
| Sep 27, 2025 | Run gel for purified BLIP-I, BLIP-II DNA, restriction digestion of BLIP-I, BLIP-II, and pSB1C3 | A | 5 | 10–11 |
| Miniprep & Glycerol Stock Preparation | B | 5 | 11 | |
| Sep 30, 2025 | Gel extraction of digested product (new method) and Ligation | A | 6 | 1 |
| Resuspension & Heat Shock Transformation of BBa_J435300 (Well O9) | B | 6 | 2 | |
| Oct 1, 2025 | Test OD for transformation products, Resuspend primers, PCR for BLIP-II | B | 6 | 3 |
| Transformation of BLIP-I and BLIP-II into pSB1C3 (DH5α and S17-1) | B | 6 | 4 | |
| Oct 2, 2025 | PCR for BLIP-I and BLIP-II; Growth of Single Colony DH5α | B | 6 | 5 |
| Oct 3, 2025 | PCR of BLIP-I and Gel run of PCR products | A | 6 | 6 |
| Subculture of S17-1 and DH5α + BLIP-I (broth) | B | 6 | 7 | |
| Oct 4, 2025 | Preparation of Streptomycin LB Broth & Competent S17-1 Cells | B | 6 | 8 |
| Transformation of pSB1C3 + BLIP-I / BLIP-II into S17-1 | B | 6 | 8 | |
| Miniprep of pSB1C3 + BLIP-I, BLIP-II, and BBa_J435320 from DH5α | B | 6 | 9 | |
| Oct 5, 2025 | Overnight Culture of BLIP-I and BLIP-II | A | 6 | 10 |
| Oct 6, 2025 | Conjugation of S17-1 (BLIP-I & BLIP-II) with DH5α | A&B | 7 | 1-2 |
| Oct 7, 2025 | Collected data on solid and liquid conjugation | A&B | 7 | 2-3 |
SEPTEMBER