Practical 4: Isolation of DNA from yeast

Results

Part B

1) Preparation of 1:50 dilution.

Initial Volume = 0.1 mL

Final Volume = 5.0 mL

2) Absorbance at 260 nm, A260 = 0.227

3) Absorbance at 280 nm, A280 = 0.128

4) % purity of DNA = (A260/A280) x 100

= (0.227/0.128) x 100

= 177 %

Questions

1. What is the concentration of DNA in your purified DNA solution (step 6)?

DNA concentration (original) = 50µg/mL x A260 x dilution factor

= 50µg/mL x A260 x (Final Volume/ Initial Volume)

= 50µg/mL x 0.227 x (5.0/0.1)

= 567.5 µg/mL

The concentration of purified DNA solution is 567.5 µg/mL.

2. What is the concentration of protein in the solution?

Protein concentration = 1.45A280 – 0.74A260

= 1.45(0.128) – 0.74(0.227)

= 1.762 x 10-2 mg/mL

The concentration of protein is 1.762 x 10-2 mg/mL.