Reagents for 1.1

10mM NaOH

MW = ~40

10mM = 0.01M = 0.4g/L

Need:
- 25 tubes of 10 mL each = 250 mL NaOH

• 13 tubes of 2 mL each = 26 mL fluorescein A

• 13 tubes of 2 mL each = 26 mL fluorescein B

= total of 300 mL NaOH needed

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)

Make A & B slightly different.

(Different enough so that they are different by the t-test. Make one above and one below the “actual” value.)

Be sure to measure a blank. In 2012, the blank was ~0.25, so Abs readings should be ~0.75