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
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 green, tubulin 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)

IF demo

IF demo kdorfman Fri, 09/27/2024 - 14:41

2024/09/30 Monday

  • Set up camera to project
  • Use left over fixed MEF
  • Set up beaker of PBS-Tw-Az
  • Materials for humid chamber
  • forceps