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 June 19 2014

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Eric Kofoid eckofoid at ucdavis.edu