TJ-Lab Summer Training held as schedule from 1 to 4 August 2023. The training was conducted online and offline simultaneously. A total of about 150 people from our laboratory and other cooperative laboratories participated in the training.
The contents of this training include molecular cloning, protein expression system and protein purification, interact expression system, and cell experiment, structural biology technology, software, scientific writing.
This training systematically introduced the theoretical knowledge and experimental operation of biochemistry and molecular biology to the freshmen, so that they had a further understanding of the scientific research life they were about to start. For senior students, they gain new insights through reviewing old material.
The training session was lead by professor Jin and organized by Dr. Zeng and ended with a group photo. We thank all participants for their contributions.
Time | Knowledge section | Topics | Presenter | Time | |
Day 1 (1st August): | |||||
a.m. | 9:00-10:00 | 1. How to do research | How to do research | 金老师 | 60mins |
10:00-10:15 | Laboratory convention | 冯喻宁 | 15mins | ||
10:15-10:25 | Vectors and cells | 赵丹 | 10mins | ||
10:25-10:40 | Tea Break | ||||
10:40-11.05 | 2. Molecular Cloning | Molecular cloning | 陈时 | 25mins | |
11:05-11:20 | E. coli competent cells preparation | 姚程炳 | 15mins | ||
11:20-11:35 | Mammalian cell culture | 刘木子樱 | 15mins | ||
午休 | |||||
p.m. | 14:30-14:45 | 3. Protein expression system and protein purification | E. coli protein expression system | 黄语洛 | 15mins |
14:45-15:05 | Yeast expression system | 姚程炳 | 20mins | ||
15:05-15:25 | Insect expression system | 姚靖雪 | 20mins | ||
15:25-15:45 | Mammalian protein expression system | 杨芸茹 | 20mins | ||
15:45-16:00 | Tea Break | ||||
16:00-16:20 | AKTA FPLC machine/demo | 曾威红 | 20mins | ||
16:20-16:50 | Columns and maintenance, Metal chelating chromatography, gel filtration, HIC, IEC concentration, preservation | 胡婧 | 30mins | ||
16:50-17:05 | Expression of TEV protease and usage, avi tag and birA(biotin) | 邓莎莎 | 15mins | ||
17:05-17:25 | MHC preparation and activity identification | 洪梦雨 | 20mins | ||
17:25-18:00 | 实验室成员大合影 | ||||
Day 2 (2nd August): | |||||
a.m. | 9:00-9:10 | 3. Protein expression system and protein purification | Large-scale plasmid extraction | 赵丹 | 10mins |
9:10-9:25 | Purification of Fab, Fc-tagged protein, Protein A/G | 朱芳 | 15mins | ||
9:25-9:45 | Theory and Application of Analytical Ultracentrifugation (AUC) | 倪康 | 20mins | ||
9:45-10:00 | Purification of antibodies in serum | 曾威红 | 15mins | ||
10:05-10:20 | Tea Break | ||||
10:20-11:00 | 4. Screening techniques for screening molecules that interact with proteins | Nanobody panning, preparation and characterization | 黄晓雪 | 40mins | |
11:00-11:15 | Nanobody panning by paramagnetic particle method | 朱利霞 | 15mins | ||
11:15-11:35 | Flow cytometry | 刘木子樱 | 20mins | ||
午休 | |||||
p.m. | 14:30-15:10 | 5. Cell expriments | Lentivirus packaging + Construction of stable cell line + Gene knockout | 陈时 | 40mins |
15:10-15:40 | Pseudovirus construction + AAV virus packaging and quality control | 姚靖雪 | 30mins | ||
15:40-16:00 | Single cell sequencing (library construction and standard analysis) | 吴齐琴 | 20mins | ||
16:00-16:15 | Tea Break | ||||
16:15-16:35 | 6. Detection technology of interaction between protein and its interaction molecule | Evaluation of vaccine efficacy | 杨芸茹 | 20mins | |
16:35-16:50 | Y2H (video) | 陆曼晴 | 15 mins | ||
16:50-17:05 | BiFc (video) | 陆曼晴 | 15 mins | ||
17:05-17:20 | ELISA and competitive ELISA | 姚靖雪 | 15 mins | ||
17:20-17:35 | Crosslink | 邓莎莎 | 15mins | ||
Day 3 (3rd August): | |||||
a.m. | 9:00-9:30 | 6. Detection technology of interaction between protein and its interaction molecule | Dot Blot, Co-IP and Western Blotting (video) | 朱利霞 | 30mins |
9:30-9:45 | EMSA | 黄晓雪 | 15mins | ||
9:45-10:00 | FP | 15mins | |||
10:00-10:30 | SPR + ITC (video) | 曾威红 | 30mins | ||
10:30-10:45 | Tea Break | ||||
10:45-11:00 | DSF (video) | 杜宇 | 15mins | ||
11:00-11:15 | 7. Application of data management software | Prism /Lasergene | 15mins | ||
11:15-11:30 | 8.Detection technology of RNA or protein expression | RNA scope | 陈琪 | 15mins | |
午休 | |||||
p.m. | 14:30-14:45 | 8.Detection technology of RNA or protein expression | Immunohistochemical (IHC) | 陶娟 | 15mins |
14:45-15:00 | 9. Characterization technology of biological macromolecular structure | CD Spectrum | 胡婧 | 15mins | |
15:00-15:15 | SAXS | 曾威红 | 15mins | ||
15:15-15:45 | 10. Principle of Cryo-EM, sample preparation, data collection and single particle selection | Cryo-EM | 王美华 | 30mins | |
15:45-16:15 | 11. Crystallization experiment | X-ray crystallography theory | 金老师 | 30mins | |
Sample preparation,Crystallization methods | |||||
16:15-16:25 | Tea Break | ||||
16:25-16:35 | Crystallization kits | 刘梦瑶 | 10mins | ||
16:35-16:55 | Optimization and salt crystals | 王美华 | 20mins | ||
Cryocrystallography,X-ray diffraction data collection | 20mins | ||||
16:55-17:15 | 12. Structure determination | Molecular graphics with Pymol | 20mins | ||
17:15-17:35 | Data processing and evaluation (HKL2000, XDS) (linux-computer demo) | 金老师 | 20mins | ||
Day 4 (4th August): | |||||
a.m. | 9:00-9:20 | 12. Structure determination | Phenix, CCP4 | 郑培义 | 20mins |
9:20-9:40 | Structural refinement | 10mins | |||
Structure valildation and deposition | 10mins | ||||
9:40-10:10 | 13. Bioinfomatics | Discover Studio | Zunera | 15mins | |
MOE | 15mins | ||||
a.m. | 10:10-10:25 | Tea Break | |||
10:25-10:45 | Molecular simulation, GROMACS | Zunera | 20mins | ||
10:45-11:00 | Docking w/ Autodock, Chimera | 15mins | |||
11:00-11:25 | Linux operation system and alphaFold | 程庆宇 | 25mins | ||
午休 | |||||
p.m. | 14:30-14:50 | 14. Animal models | Construction of mouse tumor model and validation of antibody function | 赵丹 | 20mins |
14:50-15:10 | 15. Writing | How to make a Word/PPT | 邹家莉 | 20mins | |
15:10-15:30 | Endnote/PS | 张雅馨 | 20mins | ||
15:30-15:40 | Tea Break | ||||
15:40-16:40 | Scientific writing | 金老师 | 60mins | ||
16:40-17:00 | Statistical analysis | 程庆宇 | 20mins | ||
17:00-17:10 | Summary | 金老师 | 10mins | ||
17:10--- | 聚餐 | ||||
