2024 Maresca

Slight modification of 2022

classdaydatetopics
1W9/4Tom: Intro to the course
Kate: intro to safetyfluorescence microscope
live GFP-tubulin S2 cell imaging on 8 conA coverslip dishes (prepared just before class)
pipetting part 1
2M9/9pipetting part 2
microscopy HeLa slides
S2 culture (12 flasks)
3W9/11Transformation
4M9/16mini preppcr
5W9/180.9% gel, band extraction
6M9/23invitro transcription kitconA dishes
7W9/25gel, + Nanodrop
8M9/30ds RNA treatment
IF demo (Kate)
9W10/216 conA dishes (Tom)
5 mL S2 aliquots
10M10/7immunostaining
11W10/9repeat ds RNA treatment
make 1 conA coverslip per student
12Tu10/15(M sched) IF (students seed coverslips and fix) on control and dsRNA treated cells
13W10/16looking at coverslips (Ctrl vs treated)
14M10/21presentations (in ILC?)
15W10/23transformations (PXGFP); STLC (dishes); splitting demo
16M10/28Miniprep, nanodrop, restriction digest (Bsb1)
17W10/30gel, 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
18M11/04mini prep, (skip nanodrop)
1 uL 30 min test digest (BsmB1)
45 min test gel
split cells during either digest or gel, depending on lecture
19W11/06transfection/a> of HeLa cells
20W11/13look at cells, notice transformation efficiencies
21M11/18Split parental flask
Nucleofect with Tm109 & Tm122 (1 µg each):
1 drop to dish, rest to flask
22W11/20Check nucleofected dish for GFP (take images)
Set up coverslips from flask
23M11/25IF w/Rabbit anti-Ki67
goat anti-rabbit green,
rat anti -tubulin
goat anti-rat red
THANKSGIVING
24M12/01Treat with nocodazole 1st thing
IF coverslips
26W12/04
27M12/09Poster presentations in ILC (RESERVE ROOM)