Photographic detection of fluorescent-labelled oligodeoxynucleotide in the blotting format by peroxyoxalate chemiluminescence

✍ Scribed by Kuroda, Naotaka ;Hosoki, Sayuri ;Nakashima, Kenichiro ;Akiyama, Shuzo ;Givens, Richard S.

• Publisher: John Wiley and Sons
• Year: 1998
• Weight: 380 KB
• Volume: 13
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/(sici)1099-1271(199803/04)13:2<101::aid-bio477>3.0.co;2-x

✦ Synopsis

The preparation of a fluorescent labelled oligonucleotide and its photographic detection by peroxyoxalate chemiluminescence (PO-CL) are described. Fluorescent labelling of an oligonucleotide (15-mer) was performed with naphthalene-2,3-dicarboxaldehyde to give an N-substituted 1-cyanobenz[f]isoindole (CBI) derivative (CBI-15-mer). For the photographic detection of CBI-15-mer, the bis(2,6-difluorophenyl) oxalate (DFPO)-dimethyl phthalate (DMP) system was selected to obtain a long-lived CL emission. After optimizing the conditions for the CL reaction, the system was applied to the photographic detection, and as little as 250 fmol per spot of CBI-15-mer on a membrane were detected as a visible spot with an instant photographic film.