!2022 (Maresca)

Submitted by kdorfman on Fri, 06/03/2022 - 18:13

Tom Maresca's first run of G&GA

class day date topics
1 W 9/7 Tom: Intro to the course
Kate: intro to safety, fluorescence microscope
live cell imaging on conA coverslip dishes
2 M 9/12 pipetting, S2 culture, microscopy
3 W 9/14 Transformation
4 M 9/19 mini prep, pcr
5 W 9/21 0.9% gel, band extraction
6 M 9/26 invitro transcription kit; conA dishes
7 W 9/28 gel, + Nanodrop
8 M 10/3 ds RNA treatment
9 W 10/5 16 conA dishes (Tom)
5 mL S2 aliquots
10 W 10/12 immunostaining
11 M 10/17 repeat ds RNA treatment
make 1 conA coverslip per student
12 W 10/19 IF (students seed coverslips and fix) on control and dsRNA treated cells
13 M 10/24 looking at coverslips (Ctrl vs treated)
14 W 10/26 presentations (in ILC?)
15 M 10/31 transformations (PXGFP); STLC (dishes); splitting demo
16 W 11/02 Miniprep, nanodrop, restriction digest (Bsb1)
17 M 11/07 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 W 11/09 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 M 11/14 transfection of HeLa cells
20 W 11/16 look at cells, notice transformation efficiencies
21 M 11/21 immunostaining w/ Rabbit anti-K167 (Tom to bring - what concentration?)(red secondary) & Rat anti-alpha Tubulin (Green secondary)