LUEGO: a cost and time saving gel shift procedure.
Résumé
We developed a new approach to prepare DNA probes for electrophoretic mobility gel shift assays that presents a number of advantages compared with the classical approach. The method relies on two complementary oligonucleotides containing the desired transcription factor binding sequence, one of them being extended at its 3' end with a universal tail that complements a third, short oligonucleotide labeled with a fluorophore or any other label. The use of this third "universal" oligonucleotide allows labeling many different probes with minimal time, effort and cost. We show that probes prepared this way are as effective and reliable as probes prepared by conventional methods. We refer to this short oligonucleotide as LUEGO for labeled universal electrophoretic gel shift oligonucleotide.