1.2

1.2 kdorfman Mon, 08/19/2013 - 17:18

Absorbance of Fluorescein

Per pair:

  • Black 96 well plate (not necessarily sterile, but clean)
  • 1 mM fluorescein (~ 1 mL)
  • 10 mM NaOH (~ 4.5 mL)
  • 2 2-mL tubes to make 2 mL of 10µM fluorescein (each partner does one)

Computers and Plate Readers

  • Check program Repeatability (Abs 485 nm; layout A1-B12, volume 150 µL)
  • Check program FL-Abs-by-2 (Abs 485 nm; layout C1-2, D1-12, volume 150 µL)
  • Wahoo folders must be named Tues01, Tues02, ...Tues12 and Thur01, Thur02 ... Thur12, even though the user names in the Optima program are Tue01 and Thu01, etc.
  • Import program to groups
  • Check network connections (in “my computer” connect the Z drive to Wahoo as kdorfman, kd@hh3)

solutions

solutions kdorfman Fri, 09/06/2013 - 18:47

1 mM Fluorescein

in 10 mM NaOH

It fades with time, so check the concentration of the old stocks.

MW = 332

1mM = 0.332g/L = 0.033g/100 mL

A = Ecl

Molar extinction coefficient of fluorescein at 485nm = 50358/mol.cm

so Absorbance 10^-5M = 0.50358

(1/100 of a 1mM solution =10^-5M)


10mM NaOH

MW = ~40

10mM = 0.01M = 0.4g/L