A method of one-step enzyme labelling of short oligonucleotide probes for filter hybridisation.
AUTOR(ES)
Guerasimova, A
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=148235Documentos Relacionados
- One-step labelling of oligonucleotides with fluoresceine during automated synthesis.
- Pyrosequencing™ : A one-step method for high resolution HLA typing
- A one-step method for in vitro production of tRNA transcripts
- One-step affinity purification protocol for human telomerase.
- A rapid and reliable one-step method for isolating DNA fragments from agarose gels.