This is a cached page for the URL (http://rothlab.ucdavis.edu/protocols/Z01.html). To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
DNA Quantification - TCA Precipitation

Jeff Lawrence (Z01)


1. Dilute radioactive sample to a 100 ml volume

2. Spot 5 ml of the sample onto the center of a 2.4 cm Whatman GF/C glass-fiber disc.

3. Mix 5 ml of the sample with 100 ml Salmon sperm DNA (50 mg in 20 mM EDTA).

4. Add 5 mL ice cold 10% Trichloroacetic acid (TCA). Mix well and incubate on ice for 15 min.

5. Filter the solution through a separate GF/C glass-fiber disc.

6. Wash the filter 6 times with 5 mL ice-cold 10% TCA.

7. Wash filter once with 5 mL 95% Ethanol.

8. Dry both filter under a heat lamp.

9. When dry, count each in a scintillation counter.

10. The first filter measures total radioactivity. The second filter measures radioactivity incorporated into DNA fragments greater than 20 nucleotides in length.