DNA Reaction - Radiolabeling by Random Priming

Jeff Lawrence

This protocol employs the Boeringer Manneheim Random Priming Kit.

1. Denature 25 ng duplex DNA in 9 µL ddH2O by boiling for 3 min. Immediately place on ice.

2. Add 3 µL of dGTP, dCTP, dTTP Nucleotide mix.

3. Add 2 µL Reaction Mix.

4. Add 1 µL Klenow.

5. Add 5 µL a-32P-dATP. Mix well with the pipet tip.

6. Incubate at 37° for 30 min.

7. Incubate at 95° for 2 min to denature DNA.

8. Add to hybridization bag or, optionally, remove excess labeled dATP.

Last Update: Thursday, 19-Jun-2014 11:50:20 PDT
This page has been viewed 415 times.
Eric Kofoid eckofoid at ucdavis.edu