2024 Gene & Genome (Maresca)
2024 Gene & Genome (Maresca) kdorfman Mon, 08/19/2024 - 17:30Slight modification of 2022
| class | day | date | topics |
|---|---|---|---|
| 1 | W | 9/4 | Tom: Intro to the course Kate: intro to safety, fluorescence microscope |
| live GFP-tubulin S2 cell imaging on 8 conA coverslip dishes (prepared just before class) | |||
| pipetting part 1 | |||
| 2 | M | 9/9 | pipetting part 2 |
| microscopy HeLa slides | |||
| S2 culture (12 flasks) | |||
| 3 | W | 9/11 | Transformation |
| 4 | M | 9/16 | mini prep, pcr |
| 5 | W | 9/18 | 0.9% gel, band extraction |
| 6 | M | 9/23 | invitro transcription kit; conA dishes |
| 7 | W | 9/25 | gel, + Nanodrop |
| 8 | M | 9/30 | ds RNA treatment IF demo (Kate) |
| 9 | W | 10/2 | 16 conA dishes (Tom) 5 mL S2 aliquots |
| 10 | M | 10/7 | immunostaining |
| 11 | W | 10/9 | repeat ds RNA treatment make 1 conA coverslip per student |
| 12 | Tu | 10/15 | (M sched) IF (students seed coverslips and fix) on control and dsRNA treated cells |
| 13 | W | 10/16 | looking at coverslips (Ctrl vs treated) |
| 14 | M | 10/21 | presentations (in ILC?) |
| 15 | W | 10/23 | transformations (PXGFP); STLC (dishes); splitting demo |
| 16 | M | 10/28 | Miniprep, nanodrop, restriction digest (Bsb1) |
| 17 | W | 10/30 | gel, gel extraction (1 per student), anneal primers (1 per Group), isothermal reaction, Transformation of Competent Cells 45 min gel rather than 60 min? 15 min, not 15 sec on thermocycler for isothermal rxn split HeLas while gels run |
| 18 | M | 11/04 | mini prep, (skip nanodrop) 1 uL 30 min test digest (BsmB1) 45 min test gel split cells during either digest or gel, depending on lecture |
| 19 | W | 11/06 | transfection/a> of HeLa cells |
| 20 | W | 11/13 | look at cells, notice transformation efficiencies |
| 21 | M | 11/18 | Split parental flask |
| Nucleofect with Tm109 & Tm122 (1 µg each): | |||
| 1 drop to dish, rest to flask | |||
| 22 | W | 11/20 | Check nucleofected dish for GFP (take images) |
| Set up coverslips from flask | |||
| 23 | M | 11/25 | IF w/Rabbit anti-Ki67 |
| goat anti-rabbit green, | |||
| rat anti -tubulin | |||
| goat anti-rat red | |||
| THANKSGIVING | |||
| 24 | M | 12/01 | Treat with nocodazole 1st thing |
| IF coverslips | |||
| 26 | W | 12/04 | |
| 27 | M | 12/09 | Poster presentations in ILC (RESERVE ROOM) |
HeLa for 2nd day G&GA
HeLa for 2nd day G&GA kdorfman Fri, 08/23/2024 - 16:47IF demo
IF demo kdorfman Fri, 09/27/2024 - 14:41- Set up camera to project
- Use left over fixed MEF
- Set up beaker of PBS-Tw-Az
- Materials for humid chamber
- forceps