Using Beer Lambert Law You prepared a sample of plasmid in a

Using Beer- Lambert Law:

You prepared a sample of plasmid in a total volume of 10 mL. You take 1 mL from your preparation into 100 mL of a TE buffer and determine the OD to be 0.8.

What is the total amount of plasmid you have prepared?

Solution

My primary volume or mixture is 10 mL. Now I have diluted it into 100mL TE buffter. So the dilution ratio is 1: 100.

The main concentraion in TE buffter will be 10 microgram/mL. Using Beer Lambert\'s law we know the standard absorbance concentration is A₂₆₀. Here the OD is 0.8, and we know that the absorbance is proportional to concentration. (From the same law we know that the ds DNA concentration is (@260nm) 50microgram))

0.8 OD X (10/3) * 50 microgram /mL = 133.33 microgram / ml.