1.2
1.2 kdorfman Mon, 08/19/2013 - 17:18Absorbance 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 - 18:471 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