gel retardation assay

Inge Depoortere inge.depoortere at
Thu Jun 10 09:44:30 EST 1999

I want to set up a gel retardation assay for NF-kappa-B. Can someone
give me a good and quick protocol for the preparation of the
radiolabeled oligonucleotide probe. I will start from a synthetic single
stranded probe (20bp) and want to have a double stranded labeled probe.
Can I use the end labeling procedure with T4 polynucleotide kinase?  How
should I make it double stranded. First label and then anneal (under
which conditions) or first anneal (how to do) with an overhang and then
label .
Thank you

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