2024 Gene & Genome (Maresca)

2024 Gene & Genome (Maresca) kdorfman Mon, 08/19/2024 - 17:30

Slight 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
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 immunostaining w/ Rabbit anti-K167 (Tom to bring - what concentration?)(red secondary) & Rat anti-alpha Tubulin (Green secondary)
22 M 11/20 nucleofect Guide 1 & Guide 2 (122), or just Guide 2 into WT HeLas.
DON'T USE gfp-tubulin HeLas for this
23 W 11/30 Checked transfections in cover-slip bottom dish. Nucleofection effiiency Nuc blue
24 M 11/25 IF for Ki67 tubulin and dapi.
seed glass bottom dish to view next meeting. treat cells with nocodazole at 10 am before class W
25 M 12/02 1st hour: live cell imaging of nocodazole treated cells[^*] with NucBlue
rest of class: work on posters
26 W 12/04
27 M 12/09 Poster presentations in ILC (RESERVE ROOM)