Analytical Chemistry p. 1347 - 1353 (1994)
Update date:2022-08-04
Topics:
Fujita, Satoshi
Momiyama, Masayoshi
Kondo, Yasumitsu
Kagiyama, Naoto
Hori, Samuel H.
Toru, Takeshi
A number of phosphorylated fluorochromes were synthesized and tested for the alkaline phosphatase-linked fluorescence assay of membrane-bound nucleic acids. 3-Hydroxy-N-2′-biphenyl-2-naphthalenecarboxamide phosphate ester (HBNP) was found to be the most suitable for the assay. Nonfluorescent HBNP is hydrolyzed by phospholipase bound to the probe DNA to the highly fluorescent 3-hydroxy-N,2′-biphenyl-2-naphthalenecarboxamide (HBN). HBN is insoluble enough in aqueous medium to precipitate on the nylon membrane. Spots containing as little as 5 fg of λ DNA can be successfully detected on the membrane with HBNP.
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