[en] This article describes the optimization of the hybridization signal obtained with biotinylated oligonucleotides. Optimal number and positions of biotin moieties on a 33-base oligonucleotide probe were determined. The quality of avidin-peroxidase conjugate and the choice of chromogenic substrate influenced detection sensitivity. A signal amplification method was also developed for avidin enzymatic conjugates. These improvements allowed the detection of less than 0.02 fmol of target DNA.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Zecchini, Vincent
Caucheteux, Danièle
Gretry, Jeau
Lemaître, Marc
Dommes, Jacques ; Université de Liège - ULiège > Département des sciences de la vie > Biologie moléculaire et biotechnologie végétales
Language :
English
Title :
Detection of Nucleic Acids in the Attomole Range Using Polybiotinylated Oligonucleotide Probes
Publication date :
August 1995
Journal title :
BioTechniques
ISSN :
0736-6205
eISSN :
1940-9818
Publisher :
Eaton Publishing Co., United States - Massachusetts