Lab 1 prep

Submitted by kdorfman on Wed, 08/25/2021 - 16:09
no. treated with conc [stock] time
10 untreated
2 VPA (valproic acid) 1 4 uM 20 uM over night
4 nocodazole 2 100 nM 1 mM (make 1uM 1st) overnight
2 trichostatin A 3 200 nM overnight
4 Cytochalasin D 4 250nM 10 mM 1 hour
2 Latrunculin 5 1 uM 1 mM 1 hour (in serum free medium!)

  1. Get VPA from Drew. 5X final conc.
    Add 0.5625 mL to each well containing 2.25 mL medium. ↩︎

  2. Add 97 uL DMSO to the 3 uL in tube to make 100 uL 1mM.
    Make 2mL 1uM in medium (2 uL 1mM + 1.998 mL medium).
    Add 0.25 mL to the 2.25 mL in each well. ↩︎

  3. TSA 10 mM from freezer.
    Make 1 mL 10 uM (1 uL TSA + 999 uL medium).
    Add 45 uL to each well. ↩︎

  4. Add 90 uL DMSO to the tube(makes 1 mM)
    Dilute this 1:1000 into medium (makes 1uM) (5X)
    Add .256 mL to each well ↩︎

  5. Mix 5 uL into 5 mL serum free medium (can use Fluorobrite)
    Replace the medium in each well with 2.5 mL of Latrunculin-medium mix ↩︎