A method of one-step enzyme labelling of short oligonucleotide probes for filter hybridisation.
Here we describe a method of labelling short oligonucleotide probes with enzyme without purification or chemical modifications. Biotinylated oligonucleotides as short as 10 nt are coupled with streptavidin-conjugated enzyme, hybridised and detected with enzyme-triggered chemiluminescence. The detection of hybridisation signal is linear for two orders of magnitude of target dilution. It is shown to be comparable in sensitivity with standard procedures and with radioactive detection. The method is quick, simple and has potential for automation of large-scale oligo-nucleotide hybridisation and multiplex sequencing.
Guerasimova
A
A
Max-Planck-Institute für Molekular Genetik, Ihnestrasse 73, 14195 Berlin-Dahlem, Germany andW. Engelhardt Institute of Molecular Biology, Vavilov strasse 34, 117984 Moscow, Russia. guerasimova@mpimg-berlin-dahlem.mpg.de
Ivanov
I
I
Lehrach
H
H
D000469:Alkaline Phosphatase; D019921:Biotinylation; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes; D012680:Sensitivity and Specificity; D019809:Streptavidin
D015345:Oligonucleotide Probes; D019809:Streptavidin; D000469:Alkaline Phosphatase