Simple method for preparation of fluor/hapten-labeled dUTP

Many projects, such ns multiplex-fluores- cence in situ hybridization (M-FI SH) karyotyping, require the use of relatively large amounts olf multiple f luor- or hapten-labeled nucleotides for the preparation of DNA probes. Such a requirement makes these experimental approaches prohibitively expensive for many researchers. The cost of such nucleotides can be reduced approxima tely 99% by purchasing the chemical precursors, fluor- or hapten succinimid yl esters and 5-(3-aminoallyl)-2'-deoxyuridine 5' triphosphate (AA-dUTP), a nd performing the simple coupling/purification described here. It is possib le to finish four to ten different fluor/hapten dUTP preparations of 2.5 mu M scale within a 24 h period The reagent cost for each preparation ranges from $33-$237per mu M, depending on the fluor/hapten. This laboratory uses such nucleotide preparations to prepare FISH probes by nick translation or PCR amplification.